The Campylobacter carrier rate of the bile was 23. 64%,14. 29%,7. 14%,6. 64%,5.88%, 5.55% and 3. 63% in the quails, cats, dogs, pigs, chickens, ducks and oxen respectively. The survival time of Campylobacter in the bile in vitro ranged from 4 to 7 weeks. The Campylobacter carrier rate of the intestinal content was higher than that of the bile in all the domestic animals and fowls. It is believed that the domestic animals and fowls may be an important source of Campylobacter infection in human beings according to our findings.

Objective To explore the relationship between tumor necrosis factor (TNF) gene polymorphisms in donors and recipients and the incidence and severity of acute graft-versus-host diseases (aGVHD) after unrelated allogeneic hematopoietic stem cell transplantation (alIo-HSCT). Methods Single nucleotide polymorphisms (SNPs) of TNFα-238 (G/A), TNFα-857 (C/T), TNFα-863 (C/A), TNFα-1031 (T/C), TNFβ + 252 (A/G) were analyzed by Multiplex SNaPshot analysis in 76 pairs of donors and recipients. Results Transplantation involving donors with TNFα-857 CC genotype resulted in a higher incidence of grade Ⅱ-Ⅳ aGVHD than donors with CT genotype (91.3% vs 8. 7% , P =0. 039). In the 23 patients with grade Ⅱ-Ⅳ aGVHD, no patients had TNFβ +252 AA genotype, 19 (82.6%) had GA genotype and 4 (17.4%) had GG genotype. There was a significant difference in the distribution pattern of the TNFβ +252 (AA, GA and GG) genotypes in these patients (P =0.03). There was no significant association of TNFα-238 (G/A), TNFα-863 (C/A) and TNFα-1031 (T/C) polymorphisms with the risk of aGVHD. Conclusion These results suggest donor TNFα-857 CC genotype is related to a higher incidence of grade Ⅱ -Ⅳ aGVHD, and patients with TNFβ +252 AA genotype have protection against the risk of grade Ⅱ -Ⅳ aGVHD.

Objective To determine the frequency of Toll-like receptor 4 (TLR4)polymorphisms Asp299Gly and Thr399Ile in a cohort of hematopoietic stem cell transplant recipients and their donors in China.Method We examined the polymorphisms in 208 peripheral blood samples collected from 104 recipients and their donors in a single center between 2007-2012 in Zhejiang Province,China,and Asp299Gly and Thr399Ile TLR4 gene polymorphisms were detected using the sample DNA amplification products direct sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.Result Both methods didn't demonstrate TLR4 polymorphisms Asp299Gly or Thr399Ile base mutation in our samples.Conclusion The TLR4 Asp299Gly and Thr399Ile polymorphisms are very rare in our part of the population of hematopoietic stem cell transplantation.

Objective: To evaluate the effectiveness for the construction of "mosquito-free village" in Xuejia Village, Pujiang County, Zhejiang Province, so as to provide the guidance for the construction of "mosquito-free village" in other rural areas. Methods: Density of adult mosquitoes in Xuejia Village was investigated using light trap method and density of larval mosquitoes was investigated using larval straw method from April to November each year. Totally 30 households of villagers were randomly selected, and their awareness rates of mosquito control knowledge, mosquito control behavior forming rates and satisfaction rates were surveyed through questionnaires. Investment during construction (from August 2016 to December 2018) and maintenance period (from 2019 to 2023) of "mosquito-free village" were investigated through data review and interviews. Long-term effects of "mosquito-free village" construction in Xuejia Village were evaluated in terms of mosquito density from 2016 to 2023, effectiveness of health education for villagers, satisfaction and investment. Results: Compared with the year 2016, the densities of adult and larval mosquitoes in Xuejia Village were significantly decreased from 2017 to 2023. The average monthly density of adult mosquitoes in 2023 decreased by 98.34%, and the average monthly 100 household index of larval mosquitoes decreased by 98.45% compared to 2016. The average monthly density of adult mosquitoes from 2019 to 2023 was less than or equal to one mosquito per light trap in a night, and the average monthly 100 household index of larval mosquitoes was less than or equal to five places per 100 households. The awareness rate of mosquito control knowledge was 93.33%, the behavior forming rate was 86.67%, and the satisfaction rate was 90.00%. By December 2023, the total investment during construction and maintenance period was 450 thousand Yuan, with an average annual investment of 60.7 thousand Yuan and average annual investment of 206.61 Yuan per household. The average annual investment during maintenance period was 36.2 thousand Yuan, and the average annual investment per household was 109.70 Yuan. Conclusion The mosquito density, effectiveness of health education for villagers and satisfaction of "mosquito-free village" in Xuejia Village all meet the evaluation criteria of "mosquito-free village", and the investment is reasonable, making it suitable for promotion to other rural areas.

OBJECTIVETo prepare a monoclonal antibody against human telomeric repeat binding factor 1 (TRF1) protein and explore its biological characteristics.

METHODSBALB/c mice were immunized with GST-TRF1(33-277) fusion protein for the preparation of monoclonal antibody by hybridoma technique. The obtained antibody was used for clinical assay by Western-blot and immunohistochemical staining.

RESULTSOne strain of hybridoma was obtained. It was confirmed by Western-blot that the antibody specifically recognized the 60 kD TRF1 protein. Immunohistochemical staining of the antibody showed that TRF1 protein located in the cytoplasm of epithelial cells and bone marrow cells.

CONCLUSIONA TRF1 monoclonal antibody, with high specificity was developed. It is useful for detection of TRF1 protein in tissue specimens.

Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Female ; Humans ; Hybridomas ; immunology ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; immunology ; Telomeric Repeat Binding Protein 1 ; analysis ; genetics ; immunology

OBJECTIVETo develop a novel vaccine by immobilizing interleukin-21 (IL-21) on the surface of MB49 cells and evaluate its effect in inducing specific cytotoxic T lymphocytes (CTLs) and antitumor immunity in a mouse model of subcutaneous metastatic bladder cancer.

METHODSSA-IL-21 was immobilized on the surface of 30% ethanol-fixed MB49 cells to prepare the cell vaccine. C57BL/6 mice with subcutaneous implantation of MB49 bladder cancer cells were randomized into 5 groups to receive treatments with IL-21/MB49 vaccine, soluble IL-21, GFP surface-modified MB49 cells, ethanol-fixed MB49 cells, or PBS. The tumor growth and CTL were examined to assess the antitumor efficacy of the vaccine.

RESULTSIL-21 surface-modified MB49 cell vaccine significantly inhibited the tumor growth and generated a long-lasting memory response (P<0.05). At the same effector-target (E:T) ratio, the specific CTLs induced by IL-21/MB49 vaccine showed the most potent cytotoxicity against MB49 cells (P<0.05).

CONCLUSIONWith the protein-anchor technique, IL-21 can be efficiently immobilized on the surface of MB49 cells to prepare IL-21/MB49 cells vaccine. The novel vaccine can maintain its biological activity and significantly enhance the cytotoxicity of CTLs against bladder cancer cells.

Amino Acid Motifs ; Animals ; Cancer Vaccines ; therapeutic use ; Cell Line, Tumor ; Female ; Immunotherapy ; Interleukins ; immunology ; Lymphocyte Activation ; Mice ; Mice, Inbred C57BL ; Neoplasm Metastasis ; Neoplasms, Experimental ; therapy ; T-Lymphocytes, Cytotoxic ; immunology ; Urinary Bladder Neoplasms ; therapy

Objective:To construct a transposon mutation library and screen new virulence genes of hypervirulent Klebsiella pneumoniae(hvKp). Methods:The transposon mutation library was constructed and treated with human serum. The changes in the abundance of the genes of library mutant strains were analyzed by Transposon sequencing (Tn-seq). Besides, KEGG (kyoto encyclopedia of genes and genomes) annotation and enrichment analysis were performed on the screened genes.Results:A total of 405 genes were screened out according to the abundance of the genes in the library treated with human serum was 20% lower than that without treated, and 351 genes, 86.7% of these genes were conserved in HS11286, NJST258_1, NTUH-K2044 and RJF293. Ten genes existed in strains NTUH-K2044 and RJF293 with high virulence, while these genes were absent in HS11286 and NJST258_1 with low virulence. The mutants with genes such as glycosyl transferase gene wzy, aggregator protein gene wzi and capsule transporter gene wza, which belong to the capsule polysaccharide gene clusters, could not be detected after serum treatment. The abundance of iron carriers gene clusters such as aerobacterin and salmonellin in each library changed less than one time. KEGG annotation results showed that most annotated genes were involved in amino acid metabolism, cofactor and vitamin metabolism, carbohydrate metabolism, etc. Conclusions:Tn-seq is a reliable method to screen functional genes. In this study, 405 candidate virulence genes of hvKp were successfully screened out, providing an experimental basis for further research on the function and regulation mechanism of new virulence genes of hvKp.

OBJECTIVETo analyze the epidemiology data on plague in five counties in Zhejiang province and to evaluate the risk of plague in theses areas.

METHODSWe selected five monitoring stations as a risk assessment (Qingyuan county, Longquan city, Yiwu city, Wencheng county, and Ruian city) in Zhejiang province where the plague epidemic more serious in the history. At least one constant site and 1-4 variable sites where plague occurred in history were selected for monitoring. We collected the five counties (cities) surveillance data of indoor rat density, indoor Rattus flavipectus density, the Xenopsylla cheopis index of rat, the Xenopsylla cheopis index of Rattus flavipectus in 1995-2014. Isolation of Yersinia pestis was conducted among 171,201 liver samples and F1 antibody were detected among 228,775 serum samples. Risk matrix, Borda count method, and Delphi approach were conducted to assess risk of the plague of five counties (cities) in Zhejiang province.

RESULTSIndoor rat density in Qingyuan county, Longquan city, Yiwu city, Wencheng county, Ruian city was 1.58%-5.50%, 1.13%-9.76%, 0.56%-3.67%, 2.83%-16.08%, 7.16%-15.96%, respectively; Indoor Rattus flavipectus density of five counties (cities) was 0.08%-2.23%, 0-2.02%, 0-0.54%, 0.71%-5.58%, 0.55%-4.92%, respectively. The Xenopsylla cheopis index of rat in Qingyuan county and Wencheng county was 0.011-0.500 and 0.015-0.227, respectively; The Xenopsylla cheopis index of Rattus flavipectus of Qingyuan county and Wencheng county was 0.119-3.412 and 0.100-1.430, respectively; Ruian City and Yiwu city cannot collected Xenopsylla cheopis, Long quan city only collected the Xenopsylla cheopis index of rat in the five years. Yersinia pestis were not isolated in five counties (cities).There were 3 Apodemus agrarius samples positive of plague F1 antibody test, in Longquan city and Yiwu city in 2005. Borda count method to assess the Longquan city, Yiwu (Borda point were both 321) plague risk was higher than three other regions; Delphi approach to evaluation five counties (cities) belong to the plague had a lower risk areas, according to the level of risk score (Pf) Longquan city and Yiwu (Pf was 0.314, 0.292, respectively) plague risk were higher than three other regions (Pf were all 0.292).

CONCLUSIONThe main host and media were lower in five key plague surveillance counties (cities) of Zhejiang province; The result of Borda count method and Delphi approach for risk assessment indicated that endogenous plague recrudescence was at lower level, but Longquan city and Yiwu city risk were higher than other counties (cities).

Animals ; Cities ; Epidemics ; Epidemiological Monitoring ; Humans ; Murinae ; Plague ; Rats ; Risk Assessment ; Yersinia pestis

With the progress of medical technology, the development of new drugs and the improvement of the therapeutic effect of graft-versus host disease in the last two decades, the outcomes of allogeneic hematopoietic stem cell transplantation (allo-HSCT) have been greatly improved. However, graft failure is still a rare but serious complication of allo-HSCT. HLA incompatibility, virus infection, elderly donor, uncontrolled primary disease, damage of bone marrow hematopoietic microenvironment, ABO blood group incompatibility, T cell depletion, reduced intensity conditioning, and low nucleated cell number are all risk factors for graft failure. In recent years, with the implementation of HLA haplo-identical hematopoietic stem cell transplantation, the role of donor-specific antibodies in graft failure has attracted attention increasingly. This article reviews the recent studies involving the mechanism, risk factors and prevention measures of graft failure in allo-HSCT.
Graft vs Host Disease ; epidemiology ; Hematopoietic Stem Cell Transplantation ; statistics & numerical data ; Humans ; Risk Factors ; Transplantation Conditioning ; Transplantation, Homologous ; standards ; statistics & numerical data

BACKGROUND:Achilles tendon adhesion after Achilles tendon injury can lead to decreased biomechanical properties,weakened healing ability,and ultrastructural changes of Achilles tendon,which further affects patients'daily life and work ability.Therefore,how to effectively deal with and prevent Achilles tendon adhesion has become a hot and difficult problem in clinical treatment. OBJECTIVE:To analyze the effects of biological amniotic membranes on postoperative Achilles tendon adhesion,biomechanics,and ultrastructural changes in rats with Achilles tendon rupture. METHODS:Sixty 6-week-old SD rats were selected to establish bilateral Achilles tendon rupture models and divided into two groups(n=30 per group)by the random number table method.In the model group,the severed end of the tendon was sutured directly.In the amniotic membrane group,the biological amniotic membrane was wrapped around the broken anastomosis and fixed by a suture.The adhesion,biomechanics,morphology,and structure of the Achilles tendon and the expression of p38 and ERK1/2 protein were evaluated 1,2,and 4 weeks after surgery. RESULTS AND CONCLUSION:(1)1 week after operation,the Achilles tendon and peritendinous tissues of the two groups were mildly edema,and the adhesion of the Achilles tendon tissues in the model group was more obvious.2 weeks after the intervention,the Achilles tendon and peritendinous tissues of the model group still had edema,and the adhesion degree between the Achilles tendon and the surrounding tissues was heavier than that of the amniotic membrane group.4 weeks after operation,there was no edema around the Achilles tendon in both groups,and the healing was well.The adhesion degree of the Achilles tendon in the amniotic membrane group was less than that in the model group.The maximum tension of Achilles tendons in the amniotic membrane group was higher than that in the model group at 2 and 4 weeks after operation(P<0.001).(2)Hematoxylin-eosin staining and transmission electron microscopy revealed that 1 week after operation,the tendon structure of rats of the two groups was disordered and the collagen fibers were sparsely arranged,in which the model group demonstrated obvious inflammatory reaction and adhesion to the Achilles tendon.Two weeks after operation,the model group still demonstrated obvious inflammatory response,adhesion of Achilles tendon,and irregular ordering of collagen fibers.The amniotic membrane group exhibited an orderly arrangement of collagen fibers and expansion of the endoplasmic reticulum of fibroblasts.At 4 weeks after operation,the collagen fibers of the Achilles tendon in the model group were thickened and disordered,and the rough endoplasmic reticulum was less in the fibroblasts,while the collagen fibers in the amniotic membrane group were ordered and thin,and the fibroblasts contained a large number of rough endoplasmic reticulum.(3)Four weeks after operation,western blot assay exhibited that the expressions of p38 and ERK1/2 protein in the Achilles tendon tissue of rats in the amniotic membrane group were lower than those in the model group(P<0.05).(4)The results confirm that the biologic amniotic membrane can promote the healing and inhibit the adhesion of Achilles tendon after the operation of the ruptured Achilles tendon,which may be associated with the regulation of the MAPK/ERK signaling pathway.