Abstract:Objective To detect the expression of hepatic function indices and autoantibodies in patients with chronic hepatitis B patients, patients with autoimmune liver disease and patients with chronic hepatitis B combined with autoimmune liver disease, and to evaluate the clinical significance of autoantibodies and hepatic function indexes in the early diagnosis of chronic hepatitis B combined with autoimmune liver disease.Methods A total of 109 healthy controls (HC), 72 patients with chronic hepatitis B (CHB), 74 patients with autoimmune liver disease (AILD), and 24 patients with chronic hepatitis B combined with autoimmune liver disease (CHB+AILD) in the Fifth People’s Hospital of Suzhou from 2013 to 2021 were enrolled in this study. Basic information and the value of admission hepatic function indexes and autoantibodies were collected for all enrolled samples, while no autoantibody test was performed for healthy volunteers. All data were processed using GraphPad Prism and SPSS software.Results There were no significant differences in age and gender among the four groups. The detection rates of anti-mitochondrial antibody M2 (AMA-M2) and anti-soluble acidic phosphorylated nuclear protein antibody (anti-SP100 antibodies) in CHB+AILD group [29.2%(7/24), 17.4%(4/23)] were significantly higher than those in CHB group [5.1%(3/59), 0(0/59)], suggesting that the detection of these two autoantibodies is helpful to the differential diagnosis of CHB and CHB+AILD. In addition, eight hepatic function indexes displayed significant differences among the four groups. The levels of total bilirubin and direct bilirubin in CHB+AILD group were significantly higher than those in CHB and AILD groups, while the levels of total protein and albumin were significantly lower than those in CHB and AILD groups. Alkaline phosphatase and glutamyltranspeptidase in AILD group and CHB+AILD group were significantly higher than those in CHB group. The logistic regression analysis showed that total bilirubin, direct bilirubin, albumin, alanine aminotransferase and alkaline phosphatase could form a promising prediction model, which was useful for clinicians in the differential diagnosis of CHB and CHB+AILD (area under the curve, AUC=0.902).Conclusion The combination of autoantibody and hepatic function index detection can be helpful for clinicians in the differential diagnosis of CHB, AILD and CHB+AILD, thus contributing to the early and correct diagnosis of CHB+AILD and providing theoretical basis for patients to obtain reasonable treatment and clinical cure earlier.

Objective:To observe the effect of enriched rehabilitation training on cognitive function, plasma mir-146a-5p microRNA precursor levels and inflammatory factors in persons with post-stroke cognitive impairment (PSCI).Methods:Fifty-eight persons with PSCI were randomly divided into an observation group and a control group, each of 29. The observation group was given enriched rehabilitation training, while the control group was provided with conventional cognitive rehabilitation training. The Montreal Cognitive Assessment Scale (MoCA), the Digit Span Test (DST), parts A and B of the Trail Making Test (TMT A-B) and the Modified Barthel Index (MBI) were used to assess the subjects′ cognitive functioning and their ability in the activities of daily living (ADL). Plasma levels of mir-146a-5p, IL-6 and TNF-α were detected before and after the treatment.Results:After treatment, the average MOCA, DST and MBI scores, as well as the average TMT A-B times had improved significantly for both groups. However, the observation group′s averages were significantly better than those of the control group on all three tests. After the treatment, the average plasma expression of miR-146a-5p had increased significantly in both groups, but the increase in the observation group was significantly greater. Plasma IL-6 and TNF-α levels were significantly lower than before the treatment, with the average TNF-α level in the observation group significantly lower than that of the control group.Conclusions:Enriched rehabilitation training can improve the cognition of stroke survivors more effectively than conventional cognitive rehabilitation training. That may be related to the up-regulation of plasma miR-146a-5p and reducing inflammation.

Penaeidin-2 (Pen-2) is an important antimicrobial peptide derived from the Pacific white shrimp, Penaeus vannamei, and possesses both antibacterial and antifungal activities. Recent studies suggest that recombinant penaeidins show similar activities to the native Pen-2 protein. Previous researches have shown that some antimicrobial peptides (AMPs) exhibit cytotoxic activity against cancer cells. To date, there have been no studies on the antitumor effects of Pen-2. This study evaluated the potential of recombinant pen-2 (rPen-2) in the selective killing of kidney cancer cell lines ACHN and A498, and its action mechanism. MTT assays found the maximal growth inhibition of HK-2, ACHN and A498 cells treated with 100 μg/mL rPen-2 at 48 h was 13.2%, 62.4%, and 70.4%, respectively. DNA-specific fluorescent dye staining showed a high percentage of apoptosis on cancer cells. Flow cytometry revealed that the apoptosis rate of HK-2, ACHN and A498 cells was 15.2%, 55.2%, and 61.5% at 48 h respectively, suggesting that rPen-2 induced higher apoptosis rate in cancer cells than in HK-2 cells. Laser confocal scanning microscopy demonstrated that the plasma membrane was the key site where rPen-2 interacted with and destroyed tumor cells. Scanning electron microscopy showed the morphologic changes of the cell membranes of kidney cancer cells treated with rPen-2. These results suggest that rPen-2 is a novel potential therapeutic agent that may be useful in treating kidney cancers.

We have developed a sensitive and rapid lateral-flow immunoassay (LFIA) for WSSV,using colloidal gold as an indicator.The fusion protein,VP (19+28),was expressed in E.coli,purified and used to prepare polyclonal antibodies.The purified anti-VP (19+28) IgG were conjugated with colloidal gold.Unconjugated anti-VP (19+28) IgG and goat anti-rabbit IgG were immobilized on nitrocellulose membranes.After assembly,three groups (5 individual animals in each group) of shrimp samples were tested which included healthy,moribund and dead shrimps.For each group,three different tissues (body juices,gills and hepatopancreas) were tested at the same time.In parallel,all the samples were also analyzed using PCR for comparison.Out of 45 samples tested,30 were detected as positive while 15 were classified as negative.The results of LFIA correlate with those obtained by the PCR analysis,indicating that these two detection methods have the same efficacy in the limited number of samples tested in this preliminary study.

Objective:To analyze the effects of espO gene knockout on the biological characteristics of enterhemorrhagic Escherichia coli (EHEC). Methods:Two-step methods mediated by the suicide plasmid pCVD442-Δ espO and plasmid pTrc99a were used to construct the espO gene-deleted strain (Δ espO) and the complemented mutant (CΔ espO), respectively. HeLa cells were infected with different EHEC strains to analyze the biological functions and lethal effects of espO gene during infection. Results:PCR, electrophoresis and gene sequencing showed that the Δ espO and CΔ espO mutants were successfully constructed. Compared with the wild-type strain, neither the Δ espO nor CΔ espO mutant showed significant difference in growth rate, indicating that the espO gene had no influence on the growth and replication of EHEC. Furthermore, EspO could activate the tumor necrosis factor receptor (TNF)-induced NF-κB signaling pathway, while the effector protein NleB could inhibit the process. EspO could not inhibit the death of HeLa cells induced by TNF or TNF-related apoptosis-inducing ligand (TRAIL) after EHEC infection. Conclusions:In this study, we successfully constructed the espO gene-deleted and complemented mutants of EHEC and preliminarily analyzed the interaction between espO gene and host cells and the effects of espO gene on cell apoptosis during infection, which provided reference for further research on the in vitro biochemical activity and in vivo pathogenic roles of EspO.

In recent years, polysaccharides have received much attention because of their high safety and good immunological activity. The study of polysaccharide in vivo process is a key scientific problem that needs to be solved for polysaccharide drug development. Some progress has been made in the field of polysaccharide pharmacokinetics and immunomodulation. However, due to the lack of both chromogenic and light-absorbing groups and the complex molecular structure of polysaccharides, the in vivo processes and immunomodulatory mechanisms of polysaccharides have been slow to be investigated. The effective combination of multiple techniques can break the bottleneck of difficult tracing and unknown immunomodulatory mechanism of polysaccharides in vivo, and promote the development and utilization of polysaccharides. In this paper, we systematically summarize the key techniques in the study of polysaccharide in vivo processes and immunomodulatory mechanisms in order to provide technical references and research ideas for the study of polysaccharide in vivo processes and immunomodulatory mechanisms.

Diathesis education and innovative ability cultivating of students are a new position in higher education.Exploring experiment was applied in medical undergraduates of grade 2004 for enhancing integrative diathesis and cultivating innovative spirit and ability of students.It was proved that application of exploring experiment could increase adequately learning fervor and interest of students.Teaching quality of microbiological experiment was enhanced accordingly.Application of exploring experiment was useful supplement and attempt of traditional teaching mode.

Quality education and innovative ability cultivation of students are a new position in higher education.Exploring exper- iment was applied in teaching of microbiological experiment for enhancing integrative diathesis and cultivating innovative spirit and ability of students.The practice has been proved that learning fervor of students was increased adequately.Unaided operation abili- ty,integrative analysis ability and innovative idea were enhanced,too.Accordingly,teaching quality of microbiological experiment was improved.

To determine HIV prevalence among blood donors in Zhejiang Province from 1993 to 2004 years, almost 6,600, 000 blood donors information were collected and analyzed. Every sample was screened for markers of HIV-1 and HIV-2 by using commercially available ELISA kits twice, and Western blot for confirmation if positive or suspicious result appeared. The results indicated that during the study period, prevalence rate of HIV infection was 0.85/1000 donors (0.085%), with the rising tendency from 1:600000 in 1995 to 1:37500 in 2004. The prevalence of HIV infection in Zhejiang donors was significantly lower than that in donors of other provinces, prevalence of HIV infection in male was higher than that in female. In recent years, the prevalence of HIV in blood donors was obviously increased, but the difference among various populations began to reduce. It is concluded that in a low HIV prevalence area like Zhejiang province where no obvious AIDS occurred, risk for the expansion of the HIV epidemic was on the increase. Screening procedure used in transfusion services nowaday is reliable, but a comprehensive approach is required to make the blood more safe.
Blood Donors ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV Infections ; prevention & control ; HIV Seroprevalence ; trends ; HIV-1 ; immunology ; HIV-2 ; immunology ; Humans ; Male ; Retrospective Studies ; Transfusion Reaction

WRKY transcription factors are novel transcriptional regulatory factors, which play an important role in regulating plant development, metabolism and other physiological processes. In this study, a new Dendrobium officinale WRKY transcription factor, designated as DoWRKY1 was cloned by using RT-PCR and RACE (GenBank Accession No. KF953910). Bioinformatic analysis demonstrated that, the full-length cDNA of DoWRKY1 was 1,704 bp. And DoWRKY1 contained a 1,629 bp open reading frame (ORF) that encoding a peptide of 542 amino acid residues. The putative DoWRKY1 protein contained two conserved WRKY domains and it belonged to the group I WRKY family protein. Yeast one-hybrid experiment showed that DoWRKY1 had transcriptional activation ability in yeast, and it could activate the expression of downstream report genes (His3 and Ade2). Semi-quantitative RT-PCR experiment showed that DoWRKY1 expressed in roots, stems, leaves and protocorm-like bodies. Real-time qRT-PCR proved that DoWRKY1 could be induced by methyl jasmonate (MeJA) and chitosan (Chitosan), and the expression level of this gene can reach the expression peak at 2 h and 1 h, respectively. These results are useful for further determination of the regulation function of this gene in secondary metabolism of D. officinale.
Cloning, Molecular ; Dendrobium ; genetics ; Gene Expression Regulation, Plant ; Plant Proteins ; genetics ; Transcription Factors ; genetics