Janus kinase 2,myeloproliferative leukemia virus and tet encogene family member2 mutations affect a variety of cytokines signal transduction pathway in BCR/ABL-negative myeloproliferative neoplasms (MPN). In the influence of mutation load,co-mutation and genetic susceptibility,these mutations can induce different MPN phenotypes,and affect the characteristics of patients,the distribution of peripheral blood cells and prognosis. But how these mutations contribute to disease initiation,development,and transformation needs further reseach.

A high productive poly ?-glutamic acid(?-PGA) strain PGA-N in a culture medium containing no L-glutamine was isolated from fermentation products.With the following identifications of colony mor-phology,physiological and biochemistry experiments,and genetics,the strain PGA-N was classified as a Bacillus licheniformis.According to the product environment,the base culture medium having no L-glutamine was simulated.In order to enhance the production of the strain PGA-N,the fermentation condi-tions,such as carbon source,nitrogen source,were optimized and the ?-glutamic acid production reached 5.16 g/L after getting the optimum formulation of this culture medium.PGA-N was mutagenized with com-bination of NTG and UV.A mutant PGA-N-C10 was screened which PGA production was increased from 5.16 g/L to 8.82 g/L.The study also investigated the effects of agitation speed on the cell biomass,?-PGA production and the ?-PGA molecular weight.The ?-PGA yield of PGA-N-C10 was as high as 11.00 g/L when the agitation speed was 400 r/min.

Objective: To introduce how to build a data platform of data collection and data management for multi-center birth cohort study by using REDCap. Methods: After the REDCap electronic data capture system was installed and set up, the electronic case report forms (eCRFs) were programmed to collect data in multi-centers. The rules of data quality control were programmed, and different levels of user right for access to data platform were assigned to the data managers and data clerks based on their role in research project. Results: With REDCap system being installed, the example project was created, and a series of eCRFs were established for each stage of preconception, pregnancy and childhood through follow-up. After intensive testing to improve and achieve a stable data platform, standardized trainings were provided to data-related team. The REDCap eCRFs were then put in use online. By assigning different user right of access to data platform, data entry can be from multi- research centers and survey sites. This REDCap data platform supported the cohort project on data collection and data management. Conclusion: The data platform established by using REDCap provides strong support to birth cohorts on data collection and data management. This example project of data platform can be applied to other epidemiological studies.

A poly-?-glutamic acid producing strain--BLN-2, was isolated from the soybean products. According to the biochemical characteristics and 16S rRNA, the strain was identified as Bacillus subtilis. Using soybeans as culture, the solid-state fermentation conditions of BLN-2 have been studied. The results showed that the optimal carbon and nitrogen sources of BLN-2 were glucose, fructose, NaNO3 and KNO3, respectively. The orthrogonal experiments showed, when the final concentration of the fructose which was added to the soybean culture was 0.5%, the glucose, NaNO3 and KNO3 final concentraion were 2.0%, the production of ?-PGA was the highest--89.05 g/kg. It is 48.42% higher than other comparable soybean medium under the same conditions.

A fusion gene CTB-PROIN, in which Proinsulin gene was fused to the 3' end of CTB gene by a hinge peptide 'GPGP', was constructed and cloned into pET-30a(+) to obtain a prokaryotic expression vector pETCPI. Subsequently the recombinant plasmid pETCPI was transformed into E. coli stain BL21 (DE3). After induced by IPTG, the expression product was analyzed by sodium dodecyl sulphate-polyacrylamide gel (15%) electrophoresis (SDS-PAGE), and its result indicated that the recombinant protein CTB-PROIN was expressed and accumulated as inclusion bodies. The recombinant CTB-PROIN protein accumulated to the level of 25% of total bacterial proteins. After inclusion bodies was denaturalized and refolded in vitro, significant assembly of monomers had occurred, and the recombinant protein represented assembled pentamers. The results of western blotting analysis also demonstrated that the fusion protein could be recognized by the anti-CT and anti-insulin antibody, respectively. In addition, the result of the CTB-PROIN-GM1 binding assay, that the protein could bind to monosialoganglioside specifically, showed it possesed biological activity in vitro. These results provided the possibility of developing a cheaper and more efficient oral vaccine for type I diabetes using such constructs.
Artificial Gene Fusion ; Cholera Toxin ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; G(M1) Ganglioside ; metabolism ; Proinsulin ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

Objective To explore characteristics of co-infection of Chlamydia pneumoniae(Cpn)and severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),and identify their effect on SARS-CoV-2-induced inflammatory response.Methods Patients with coronavirus disease 2019(COVID-19)who received treatment in a hospital in Chenzhou City from December 20,2022 to February 20,2023 were selected.According to the severity of COVID-19,severe and critical cases were classified as the severe symptom group,while mild and moderate cases were classified as the mild symptom group.Meanwhile,according to the age of patients(≥18 years old as adults,＜18 years old as juveniles),they were divided into the adult severe symptom group,adult mild symptom group,juvenile severe symptom group,and juvenile mild symptom group.Propensity score was adopted to match age,gender,and under-lying diseases of patients in severe symptom and mild symptom group in a 1∶1 ratio.Bronchoalveolar lavage fluid(BALF),throat swabs,and serum specimens of patients were collected.Cpn IgG/IgM antibody was detected by enzyme-linked immunosorbent assay(ELISA),levels of 12 common cytokines(including interleukin-8[IL-8])in BALF were detected by flow cytometry,differences among groups were compared.Results A total of 102 patients were included,with 61 severe and critical(severe symptom)patients,as well as 41 mild and moderate(mild symp-tom)patients.There were 71 patients aged ≥18 years and 31 juvenile patients aged＜18 years.There were 39 pa-tients in the adult severe symptom group and 32 in the adult mild symptom group,and 30 pairs were successfully matched through propensity score analysis.There were 22 patients in the juvenile severe symptom group and 9 in the juvenile mild symptom group,and 8 pairs were successfully matched through propensity score analysis.Among COVID-19 patients,the positive rates of Cpn IgG and IgM were 36.27％(n=37)and 8.82％(n=9),respective-ly,with 1 case positive for both Cpn IgG and IgM.The level of interferon(IFN)-α in serum specimens from adult patients with severe symptom combined with positive Cpn IgG was higher than that of IgG negative patients(P=0.037).There was no statistically significant difference in the levels of other cytokines in BALF and serum speci-mens between the two groups of patients(all P＞0.05).The levels of IL-8 and IL-17 in serum specimens of patients with positive Cpn IgG in the adult mild symptom group were both higher than those in Cpn IgG negative patients(both P＜0.05).The levels of IL-8 in both BALF and serum specimens from Cpn IgM positivity patients in the ju-venile mild symptom group were higher than those from patients with negative Cpn IgM(both P＜0.05).Logistic regression analysis results showed that Cpn IgG and IgM positivity were not risk factors for the development of se-vere COVID-19.Conclusion Combined Cpn infection is not a risk factor for the development of severe symptom in COVID-19 patients,and Cpn infection has limited impact on the secretion of inflammatory factors caused by SARS-CoV-2.

Infra-slow oscillation (ISO) is a kind of brain rhythm between 0.01 and 0.5 Hz. ISO is widely distributed in multiple brain regions. As an important psychophysiological activity, the ISO interacts with high-frequency neural rhythm via cross-frequency coupling, but has different activity patterns from high-frequency neural activity. Physiologically, the ISO may be generated by the dynamic activity of thalamus, glia, and ions. Psychologically, the frequency, amplitude, and phase of ISO could all regulate cognitive activities, but in different ways. Investigations on the ISO expands the neural rhythm research to lower frequency range, further promoting the construction of rhythmic theory of brain function.
Brain ; Thalamus