Objective To investigate the effect of glutamine intensified TPN on nutritional state and immunofunction in the postoperative patients with gastrointestinal tract tumour.Methods 20 patients were divided into two groups including standard TPN group (control group,10 cases) and Glutamine intensified TPN group(study group,10 cases). Period of treatment was 8 days.Immunofunction(IgG,IgM,IgA,C 3,C 4), serum albumin , nitrogen balance were tested.Results (1)The patients serum albumin in the study group was increased significantly as compared with control group (P

To rapidly select potent anti-VSTM1-v2 scFv (single-chain antibody fragment) by construction and screening of a humanized scFv library in which a murine VH-CDR3 library was grafted onto a human scFv framework. A murine VH-CDR3 library was amplified from anti-VSTM1-v2 murine cDNA and grafted on human scFv (VH3-VK1) framework. Anti-VSTM1-v2 scFv templates were selected and enriched through ribosome display, TA-cloned into expression vector, and transformed into BL21 (DE3) for soluble expression of target scFv. A total of 1000 clones were randomly picked. Positive ones were first identified using colony PCR, indirect ELISA, Western blotting and then verified with sequencing and dose response ELISA. At last an anti-VSTM1-v2 humanized scFv with good binding affinity (EC50 = 21.35 nmol x L(-1)) was selected from the humanized library of 10(12) members generated in this study. This scFv antibody might have potential applications. This study provides a new approach for rapid screening of humanized antibodies.

Total mRNA was extracted from lymphocytes separated from the peripheral blood of allergic patients, and then variable region of heavy chain (VH) and variable region of light chain (VL) cDNA library were constructed by RT-PCR. Human scFv templates for rabbit reticulocyte lysate ribosome display were assembled by primers and linker peptide (Gly4Ser)3. mRNA bound in antibody-ribosome-mRNA complexes was recovered using in-situ single primer RT-PCR, and three rounds of anti-IgE scFv DNA were enriched. The target DNA fragments were double enzyme digested and ligated into plasmid pET22b (+), followed by transformation in E. coli Rosseta (DE3). Positive clones were screened using clone PCR, Dot blotting and antigen ELISA. The correct lengths of VH (400 bp) and VL (710 bp) PCR products were obtained. The expected 1,000 bp ribosome display templates were also observed in agarose gel electrophoresis. After three rounds of ribosome display target sequences were effectively enriched, leading to a library of 10(13) members. Antibodies with the highest ELISA value for IgE were generated in the strain pET-IgE-6. A human anti-IgE scFv library was successfully constructed as described herein. Ribosome display using single primer in-situ RT-PCR as the recovery procedure effectively enriched target sequences. Anti-IgE scFv with high affinity and specificity were identified. The prepared human anti-IgE scFv fragment might be self-developed to a lead drug for treating asthma. Our study provides an alternative method for rapid discovery of human antibodies of therapeutic importance.

BACKGROUND:With the wide application of bone repair scaffold in the field of medicine, nano-hydroxyapatite (nHA) and silk fibroin (SF) both of which have good biological properties have become research hotspots in recent years.OBJECTIVE:To study the feasibility of nHA/SF composite materials loaded with recombinant human bone morphogenetic protein 2 (rhBMP-2) to restore the initial stability of the spinal segment in rabbits.METHODS: Thirty-six male health New Zealand rabbits were randomly divided into three groups, followed by preparation of spinal instability models. Autogenous iliac bone, nHA/SF composite, and rhBMP-2/nHA/SF composite were implanted into the L4/5 spinal segment in autologous bone group, nHA/SF group and rhBMP-2/nHA/SF group, respectively. X-ray exmaination was performed at 12 weeks postoperatively, and then the animals were killed for gross observation. The stability of the fusion segments was tested through a tensile machine. Histologically, bone graft fusion at the surgical site was observed.RESULTS AND CONCLUSION:(1) Findings from the gross specimen observation showed that the specimens at the fusion site presented with a hard texture. Obvious signs of fusion were visible in the autologous bone group, followed by the rhBMP-2/nHA/SF group, while no signs of fusion were detected in the nHA/SF group. (2) At 12 weeks postoperatively, a large number of trabecular bones grew into the boundary between the vertebral body and the iliac crest graft block in the autologous bone group. A little trabecular bone was found in the boundary in the nHA/SF group, while a lot of trabecular bone tissues were found in the boundary in the rhBMP-2/nHA/SF group. In accordance with the standard of fusion, there were 10, 3, and 9 rats in the autologous bone, nHA/SF and rhBMP-2/nHA/SF groups, respectively. (3) The range of motion of the spine in the rhBMP-2/nHA/SF showed no statistical difference from that in the autologous bone group, but was significantly higher than that in the nHA/SF group (P < 0.05). (4) Osseous connection was found around the bone graft in the autologous bone and nHA/SF groups, but no mature bone tissue was visible in the latter group. In the rhBMP-2/nHA/SF group, a large number of new capillaries was found and permeated into the spinal tissues. In summary, nHA/SF composite materials loaded with rhBMP-2 possess good biocompatibility, mechanical properties and bone induction ability, which can rebuild the initial stability of the spine in a short time.

We detected Zika virus (ZIKV) in a febrile case returning from Suriname and entry China from Guangzhou Baiyun International Airport Port.Serum and saliva samples were collected from a suspected case returning from Suriname.We detected ZIKV RNA using real-time fluorescence RT-PCR methods by both in-house reagent and commercial detection kits.RT-PCR detection was carried out with saliva sample and sequence analysis was performed.Phylogenetic tree was constructed to analyze the source of imported cases.Real-time fluorescent RT-PCR result showed that saliva was detected ZIKV RNA positive while for serum was weakly positive.A specific 1 500 bp fragment in size was amplified with saliva sample by RT-PCR.Sequence analysis showed 99％ homologous to the corresponding sequence of Brazil ZIKV (GenBank No.KX197250).Phylogenetic tree indicated it was located on African lineage.According to the epidemiological investigation results,clinical manifestations and nucleic acid detection of case,the suspected case was confirmed to infect Zika virus,being the first case from Suriname into Guangdong Province.

Objective:To develop a single-tube one-step multiplex nested real-time reverse transcription polymerase chain reaction (RT-PCR) assay for the simultaneous detection of 2019-nCoV, influenza A virus, influenza B virus and internal-control with human-derived gene.Methods:This study included 30 positive specimens for 2019-nCoV nucleic acid detection and 336 screening specimens collected from the arrivals at Guangzhou Baiyun Airport between February 2020 and February 2022. Sixty-four positive specimens of other respiratory pathogens were also collected from the arrivals at Guangzhou Baiyun Airport during the three-year period before the occurrence of COVID19 outbreak in 2020, and 7 positive viral strains of respiratory pathogens were provided by collaborative laboratories. In order to establish a set of multiplex nested real-time RT-PCR assay, a group of primers and probe combinations for a multiplex nested real-time RT-PCR was designed and screened according to a selection of nucleotide conserved regions of the ORF and N genes of 2019-nCoV and the M gene of influenza A and B viruses, while nested amplification primers and probe for the internal-control with human-derived gene were introduced. Then the prepared pseudovirus-positive quality control and sample discs were applied to evaluate the sensitivity and specificity. Clinical specimens were performed to validate the applicability of the method.Results:The results show that the established one-step multiplex nested real-time RT-PCR assay can specifically detect 2019-nCoV and influenza A and B viruses, with the limit-of-detection of about 125 copies/ml for 2019-nCoV and about 250 copies/ml for influenza A and B viruses. Totally 101 positive samples of various respiratory pathogens were detected, showing that the detection sensitivities of 2019-nCoV and influenza A and B viruses were 96.67%, 92.86% and 96.15%, respectively, with the specificity of 100%. No false-positive detection was found in the applied detection of more than 300 clinical samples.Conclusions:A one-step multiplex nested real-time RT-PCR assay for 2019-nCoV, influenza A and B viruses and human-derived gene internal-control was developed. The assay has good sensitivity and specificity and can be used for rapid screening of 2019-nCoV and influenza A and B viruses in high-volume samples.