Exploration on multifunctional linkage and scientific use of medical colleges psychological experimental teaching center based on the network background / 中华医学教育探索杂志

The experimental teaching of psychology is not only an important course in the undergraduate and graduate education of applied psychology, but also can assist the practical teaching of quite a few other subjects. It clearly divides three basic functions of experimental psychology teaching center: teaching, scientific research and service, which play an important role in facilitating teachers and students to understand corresponding courses. At the same time, it is discussed and proposed to strengthen the internal and external scientific linkage of the experimental center under the network background, and put forward the use of Internet technology, in order to improve the scientific use of the experimental teaching center, and reflect its maximum value, thereby achieving the purpose of university laboratory joint construction and serve the university to cultivate innovative and compound personnels.

Research and application progress in adoptive re-transfusion of regulatory cells in organ transplantation / 器官移植

Rejection and adverse reactions caused by long-term use of immunosuppressants severely affect the survival rate and quality of life of organ transplant recipients. Immune tolerance induction plays a key role in improving the survival rate and quality of life of organ transplant recipients. In recent years, tremendous progress has been achieved in adoptive re-transfusion of regulatory cells. In this article, research progress in regulatory T cell (Treg), myeloid-derived suppressor cell (MDSC) and regulatory B cell (Breg) in animal experiment and clinical application was reviewed, and the main clinical problems of adoptive re-transfusion of regulatory cells, the application of chimeric antigen receptor Treg and the concept of cell therapy in immune evaluation were summarized, aiming to deepen the understanding of regulatory cell therapy, promote the application of regulatory cells in immune tolerance of organ transplantation, and improve clinical efficacy of organ transplantation and the quality of life of recipients.

Establishment of a membrane metalloproteinase TRABD2A blocking method for HIV reservoir detection / 中华检验医学杂志

Objective:To investigate the therapeutic effect of humanized TRAB domain-containing protein 2A (TRABD2A) monoclonal blocking antibody to HIV-1 reservoir cells, and to explore novel methods for measuring the sizes/capacities of HIV-1 infected reservoirs in HIV-1 infected individuals on receiving combined antiretroviral therapy (cART).Methods:A total number of 51 subjects were collected from the First Affiliated Hospital of China Medical University from May 2021 to December 2021. Among them, there were 2 healthy persons, 41 HIV-1 infected persons receiving cART (cART group) and 8 HIV-1 infected persons not receiving cART (no cART group). Humanized TRABD2A monoclonal antibody was constructed based on the phage display technology, the PBMCs and CD4+T cells separated from the peripheral blood mononuclear cells (PBMCs) and CD4+T cells of HIV-1 infected patients treated with receiving cART, or the HIV-1 infected patients without cART treatment and healthy controls were treated with TRABD2A monoclonal antibodies. The luciferase reporter system, single molecule immune array detection technology and other methods were used to detect the virus content in the supernatant of cell culture. At the same time, flow cytometry and fluorescence real-time quantitative polymerase chain reaction were used to detect the activation of the treated cells and the expression of virus genes. The statistical differences between different treatment the amount of virus release and the level of surface activation markers CD25, CD69, human leukocyte antigen DR (HLA-DR) of different groups in the amount of virus release and the expression of surface activation markers CD25, CD69, HLA-DR were compared.Results:The PBMCs of HIV-1 infected persons receiving cART were tested for HIV-1 production after being treated with humanized TRABD2A monoclonal antibody. The amount of virus released by the untreated group was 0 (0, 440), and the amount of virus released by the use of negative antibody was 0 (0, 390). There was no significant difference between the two ( P>0.05). The amount of virus released by the use of positive antibody was 1 259 (0, 4 269), 3 142 (1 292, 5 060), compared with the amount of virus released by the use of negative antibody, The difference was statistically significant ( P<0.05). The healthy control PBMC was used to conduct multiple dilutions to the infected PBMC. After positive antibody treatment, the amount of virus release decreased in equal proportions [the HIV-1 production corresponding to 5, 25, 125, 625 times of undiluted, diluted PBMC was 4 670 (3 339, 7 697), 1 860 (1 509, 4 615), 1 550 (1 150, 2 680), 602 (255, 1 441), 2 (0, 37), respectively].In addition, there was no significant difference in the resting state of cells treated with TRABD2A antibodies compared with the untreated group (The percentage of CD25 positive cells in the untreated group and positive antibody 1 treated group were 3.89±1.31 and 4.60±1.74, the percentage of CD69 positive cells were 2.50±1.27 and 2.18±0.51, and the percentage of HLA-DR positive cells were 7.66±3.78 and 8.79±3.42, respectively, P>0.05). The viral gag expression levels of untreated and positive antibody 1 were 1 and 0.82±0.55, respectively, with no significant difference. Conclusions:The humanized TRABD2A monoclonal antibody can effectively block the protein activity of TRABD2A, and can significantly promote the release of progeny viruses from viral reservoir in the peripheral blood of HIV-1 infected persons without changing the cell resting state and the whole genome transcription level. The amount of virus released in this way is positively related to the number of reservoir cells.

Changes and significance of lymphocyte subsets in recipients with acute rejection after liver transplantation / 器官移植

Objective To evaluate the changes and significance of lymphocyte subsets in the recipients with acute rejection after liver transplantation. Methods The recipients presenting with acute rejection after liver transplantation were assigned into the rejection group (n=17), and their counterparts with stable liver function were allocated into the control group (n=17) according to the ratio of 1∶1 by propensity score matching method. The incidence of acute rejection after liver transplantation was analyzed, and the concentration of tacrolimus in the recipients was compared between two groups. The absolute value and proportion of lymphocyte subsets in peripheral blood were compared between two groups. The diagnostic value of lymphocyte subsets for acute rejection after liver transplantation was assessed by the receiver operating characteristic (ROC) curve. The absolute value and proportion of lymphocyte subsets in the rejection group were compared before and after treatment. Results Among 17 recipients in the rejection group, 4 cases developed acute rejection within postoperative 28 d, and 13 cases had acute rejection within postoperative 29-180 d. No significant difference was noted in the tacrolimus concentration between two groups (P=0.295). Compared with the control group, the proportions of peripheral blood T cells, CD4+T cells, B cells and natural killer (NK) T cells were significantly increased in the rejection group (all P < 0.05). The elevated proportion of NKT cells in the early stage after liver transplantation was an independent risk factor for acute rejection following liver transplantation[odds ratio (OR) 1.774, 95% confidence interval (CI) 1.059-2.971, P=0.029]. ROC curve analysis showed that the area under curve (AUC) of CD4+T cells, B cells and NKT cells was 0.76, 0.73 and 0.77, respectively. The AUC of combined use of CD4+T cells, B cells and NKT cells was 0.89, with a cut-off value of 0.69, sensitivity of 0.706 and specificity of 0.941. After corresponding treatment, all recipients were gradually recovered, and liver functions were eventually restored to normal in the rejection group. After treatment, the proportion of T cells, CD4+T cells, CD8+T cells and NK cells was significantly decreased (all P < 0.05). Conclusions The elevated proportion of NKT cells indicates an increased risk of acute rejection after liver transplantation. Combined use of CD4+T cells, B cells and NKT cells may deliver early detection and diagnosis of acute rejection after liver transplantation.

Application study on adoptive transfusion of tolerogenic dendritic cells in promoting immune tolerance of liver transplantation in rat models / 器官移植

Objective To investigate the role of tolerogenic dendritic cell (tolDC) in inducing immune tolerance in liver transplantation. Methods Liver transplantation rat models of spontaneous tolerance [Brown Norway (BN)→Lewis, tolerance group, n=6] and acute rejection (AR) (Lewis→BN) were established. In AR rat models, tolDC transfusion was performed in the study group (tolDC group, n=6) and no intervention was given in the control group (AR group, n=6). The survival time of rats in each group was observed. The transplant liver tissues of rats were prepared for pathological examination in each group. The expression of myeloid dendritic cell (mDC) and plasmacytoid dendritic cell (pDC) in rat peripheral blood, transplant liver, spleen and lymph nodes in each group was detected by flow cytometry. The expression levels of serum interleukin (IL)-10 and interferon (IFN)-γ in each group were measured by enzyme-linked immune absorbent assay. Results Pathological manifestations of rats in the AR group mainly included inflammatory cell infiltration and tissue structural disorder in transplant liver, and the survival time was 7-14 d. In the tolDC and tolerance groups, the transplant liver tissues were almost normal, and the longest survival time exceeded 100 d. Compared with the AR group, the expression levels of CD11+mDC in peripheral blood, transplant liver, spleen and lymph nodes of rats were significantly down-regulated in the tolerance and tolDC groups (all P < 0.05), and those of CD86 and major histocompatibility complex (MHC)Ⅱon the surface of CD11+mDC were also significantly down-regulated (all P < 0.05). Compared with the AR group, the expression levels of pDC in peripheral blood, transplant liver, spleen and lymph nodes of rats were significantly up-regulated in the tolerance and tolDC groups (all P < 0.05), whereas those of MHCⅡon the surface of pDC were all significantly down-regulated (all P < 0.05). Compared with the AR group, the expression levels of serum IL-10 were significantly up-regulated, and IFN-γ were significantly down-regulated in the tolerance and tolDC groups (all P < 0.05). Conclusions As tolDC subsets, mDC and pDC play a positive role in regulating the incidence of graft immune tolerance in rats after liver transplantation.

Serotype distribution and drug resistance of Shigella in children with diarrhea from 2008 to 2017 in Sui County, Henan Province / 中华传染病杂志

Objective:To explore the serotype distribution and drug resistance of Shigella in stool samples of children under five years old with diarrhea from 2008 to 2017 in Sui County, Henan Province. Methods:A total of 4 721 stool samples of children under five years old with diarrhea were collected from Doufuyuan Clinic of Sui County during 2008 to 2017, and Shigella strains were isolated through bacterial culture. The slide agglutination test was used for serotyping of Shigella strains. Two hundred of seventy-one Shigella strains were selected in proportion, and multiple gradient polymerase chain reaction was used to detect virulence genes and Kirby-Bauer agar method was used for drug sensitivity. Trend chi square test was used to analyze the annual trend of drug resistance. Results:The detection rate of Shigella strains in 4 721 fecal samples was 20.69% (977/4 721). A total of 977 Shigella strains were divided into 13 serotypes in two groups, including 77.79%(760/977) Shigella flexneri strains and 22.21%(217/977) Shigella sonnei strains.The top three serotypes detected alternately every year.The dominant gene pattern of Shigella flexneri was Shigella enterotoxin ( shET)-1+ , shET-2+ , invasion plasmid antigen H ( ipaH)+ , invasion-associated locus ( ial)+ , accounted for 84.04%(179/213) and that of Shigella sonnei was shET-1-, shET-2+ , ipaH+ , ial+ , accounted for 46.55%(27/58). The drug resistance rates of 271 Shigella strains to ampicillin, nalidixic acid and tetracycline were more than 90% and the strains were more sensitive to cefepime and ceftazidime.The drug resistance rates to cefotaxime, cefepime, ciprofloxacin, chloramphenicol and sulfamethoxazole/trimethoprim increased year by year, and all had statistically significant differences ( χ2=24.027, 7.232, 6.039, 4.764 and 6.809, respectively, all P< 0.05). There were 98.52%(267/271) strains resistant to more than three kinds of drugs. The resistance rates of Shigella flexneri to ciprofloxacin, norfloxacin, and chloramphenicol were higher than those of Shigella sonnei, and the resistance rates to gentamicin and sulfamethoxazole/trimethoprim were lower than those of Shigella sonnei. The differences were statistically significant ( χ2=31.866, 14.868, 83.036, 68.534 and 14.738, respectively, all P<0.01). Conclusions:The major serotypes of Shigella in children under five years old in Sui County are constantly changing from 2008 to 2017. The dominant gene patterns of different serotypes are different. Most isolated strains have multiple drug resistances, and different serotypes have different resistance profiles.

Analysis on the epidemiology and etiology characteristics of first imported Chikungunya fever case in Henan Province in 2017 / 中华预防医学杂志

To study the epidemiology and etiology characteristics of first imported Chikungunya fever case in Henan province, China, 2017. The patient was confirmed by Chikungunya virus (CHIKV) infected as CHIKV ribonucleotide was continuously detected in his serum specimens. BHK-21 cell line was used for virus isolation, the strain was named CHIKV/Henan001/2017. CHIKV/Henan001/2017 belonged to genotype ECSA. The highest ribonucleotide homology sequence of highly conserved region E1 with CHIKV/Henan001/2017 was hk02 strain (99.8%), who was an imported strain to Hong Kong, China, 2016. Epidemiological information and laboratory testing confirmed it was an imported Chikungunya fever case in Henan province, 2017. No secondary case has been reported.

Laboratory diagnosis and molecular tracing of dengue bordline cases in Henan Province, 2017 / 中华预防医学杂志

Objective@#To confirm the laboratory diagnosis of dengue bordline cases reported in Henan Province and trace its origin from molecular level in 2017.@*Methods@#The study samples were blood samples (3-5 ml), which came from 8 suspected cases of dengue fever reported in the 2017 direct reporting system of Henan provincial infectious disease monitoring network. Meanwhile, case investigation was conducted according to National dengue fever surveillance programme. Serum were separated from blood samples and tested for Dengue NS1 antigen, IgM & IgG antibodies, and dengue RNA. According to dengue diagnosis criteria, confirmed cases were identified by testing results. Samples carried dengue RNA performed for real-time PCR genotyping and amplification of E gene. Then, the amplicons were sequenced and homological and phylogenetic analyses were constructed.@*Results@#8 serum samples of suspected dengue cases were collected in Henan Province, 2017. Six of them were diagnosed as dengue confirmed cases. All the dengue confirmed cases belonged to outside imported cases, 5 of them were positive by dengue RNA testing. Genotyping results showed there were 1 DENV1 case, 2 DENV2 cases and 2 DENV3 cases. A DENV2 case and a DENV3 case of this study were traced its origin successfully. The sequence of Pakistan imported DENV2 case belongs to cosmopolitan genotype, which was the most consistent with Pakistan's DENV2 KJ010186 in 2013 (identity 99.0%). The sequence of Malaysia imported DENV3 case belongs to genotype I, which was the most consistent with Singapore's DENV3 KX224276 in 2014(identity 99.0%).@*Conclusion@#The laboratory diagnosis and molecular traceability of dengue cases in Henan Province in 2017 confirmed that all cases were imported and did not cause local epidemics.

Surveillance of avian influenza virus in poultry environment in Henan Province, 2013-2017 / 中华微生物学和免疫学杂志

Objective To monitor the environmental contamination with avian influenza virus (AIV) in Henan Province. Methods Environmental samples were collected every month from seven moni-toring sites in Henan from 2013 to 2017. Real-time RT-PCR method was performed to detect the nucleic acid of influenza A (Flu A), H5, H7 and H9 viruses in poultry environmental samples. Results A total of 2538 environmental samples were collected and 202 (7. 96％ ) of them were positive for Flu A nucleic acid, including 16 positive for H5 (0. 63％ ), eight positive for H7 (0. 32％ ) and 161 positive for H9 (6. 34％ ). The detection rate of Flu A increased dramatically from 2013 to 2017 except for a small fluctuation in 2015. However, H7 subtype AIV was detected only in 2015 and 2017. The highest detection rate of AIV was in February, followed by that in January. Among different environments, the highest detection rate of Flu A was in live poultry market, which was 13. 69％ , followed by that in poultry slaughtering plant (2. 58％ ) and poultry farm (0. 58％ ). The detection rates of Flu A in swab samples of poultry plucker and cutting board, stool specimens and poultry drinking water were 28. 57％ , 13. 76％ , 5. 70％ and 5. 26％ , respectively.Conclusion Contamination of H5 / H7 / H9 AIV did exist in poultry environment in Henan and was getting worse. Increasingly diversified sources and sale channels were the main causes of serious contamination of AIV. In order to effectively prevent and control human infection with AIV, live poultry in areas where human infection with AIV was confirmed should be blocked and banned to be sold to others areas.