[Current prophylaxis and treatment of venous thromboembolism in intensive care units of 41 tertiary hospitals of Zhejiang Province].

To investigate the prophylaxis and treatment of venous thromboembolism (VTE) in intensive care units (ICU) in Zhejiang Province. A cross-sectional questionnaire survey for ICUs doctors was conducted online at 41 tertiary hospitals in Zhejiang Province, which included basic information of ICUs, VTE-related installations, knowledge and monitoring system. The data collected from the questionnaire were summarized and analyzed. Most ICUs were comprehensive. The ratio of physicians to beds, ratio of nurses to beds, and the ratio of rehabilitation physicians to beds were 0.52∶1, 2.12∶1, and 0.03∶1 respectively. Thirty-five over 41 (85.4%) ICUs completed VTE risk assessment within 24 hours. Twenty-five over 35 ICUs (71.4%) chose the Caprini model to assess VTE risk. Ultrasound was preferred in 92.7% (38/41) of ICUs equipped ultrasound. Almost all (40/41, 97.6%) of hospitals possessed intermittent pneumatic compression (IPC) as mechanical prophylaxis. The ratio of IPC to bed was 0.22∶1. Low-molecular-weight heparin (92.7%, 38/41) was the initial treatment in ICUs. VTE prophylaxis system has been established in 92.7% (38/41)hospitals and 87.8% (36/41) ICUs,related personnel groups were set up in 75.6% (31/41) hospitals and 58.5% (24/41) ICUs including 68.3% (28/41) hospitals with multidisciplinary team. VTE prophylaxis system in different hospitals was heterogeneous. VTE risk assessment models in different ICUs were not identical. There were unmet clinical needs of VTE mechanical prophylaxis equipment. In conclusion, VTE screening, evaluation and prophylaxis protocols in ICUs still need to be standardized and improved.

[The prognostic role of the programmed death-1 expression on T lymphocytes in septic patients].

Objective: To investigate the value of programmed death-1（PD-1） expression on the T lymphocytes for the prognosis of septic patients. Methods: From September 2017 to May 2019, septic patients were included in Department of Intensive Care Unit at 6 hospitals. The PD-1 expression on T cells were measured by flow cytometry. Logistic regression was conducted to analyze independent risk factors related to death within 28 days，and receiver operating characteristic curve(ROC) was conducted to evaluate the prognostic value of PD-1 expression on T cells in septic patients. Results: A total of 64 septic patients were enrolled to this study，including 32 survivors and 32 deaths. The PD-1 expression on T cells in the death group was significantly higher than that in the surviving group (P<0.05). Correlation analysis showed that the percentages of PD-1+/CD3+T cells and PD-1+/CD8+T cells were positively correlated with procalciton in (r=0.313, P =0.015；r=0.375, P=0.003), logistic regression analysis showed that the percentages of PD-1+/CD3+，PD-1+/CD4+，PD-1+/CD8+T cells were independent risk factors for the death of sepsis patients. The percentage of PD-1+/CD3+T cell was 3.63%, with AUC 0.842, sensitivity to predict the mortality 96.43% and specificity 59.38%, (P<0.000 1). The percentage of PD-1+/CD4+T cell was 4.65%, with AUC 0.847, sensitivity 96.43%, specificity 62.50%，(P<0.000 1). The percentage of PD-1+/CD8+T cell was 3.91%, with AUC 0.771, sensitivity 64.29%, specificity 81.25%，(P=0.000 3). Conclusions: The T cell PD-1 expression is an independent risk factor to predict the 28-day mortality in septic patients. Combining the proportions of PD-1+/CD3+, PD-1+/CD4+and PD-1+/CD8+T cells may further enhance the predictive value for death.

[Antimicrobial resistance changes of carbapenem-resistant Enterobacteriaceae strains isolated from children].

Objective: To investigate the prevalence and resistance changes of carbapenem-resistant Enterobacteriaceae (CRE) strains isolated from children patients of Chinese Bacterial Resistance Surveillance Network (CHINET) from 2005 to 2017. Methods: Antimicrobial susceptibility testing was carried out by disk diffusion method (KB method) and automated systems. Results were analyzed according to the Clinical and Laboratory Standards Institute (CLSI) 2017 edition standards. Results: Among the 4 481 CRE clinical strains, the overall prevalence of CRE in children was 6.4%, including 8.8% in neonatal period, 7.3% in infancy, 3.8% in early childhood, 4.0% in preschool, 4.7% at school age and 7.4% of puberty. The CRE prevalence of citrobacter spp. remained stable in 2005-2017, whereas other bacteria showed an upward trend, which was higher than that of the adult group (P<0.01). Among the 4 481 CRE strains, there were 2 905 strains of Klebsiella spp. (64.8%), 813 strains of Escherichia coli (18.1%), 549 strains of Enterobacter spp.(12.3%), and 65 strains of Citrobacter spp.(1.5%). Among the 4 481 CRE strains, 20.7%, 13.3%, and 11.8% were from the intensive care unit (ICU), neonatal department and internal medicine wards, respectively. Specimens were distributed as respiratory (42.8%), urine (26.3%), and blood (14.9%). The results of antimicrobial susceptibility testing exhibited that the CRE strains were highly resistant to most commonly used antimicrobial agents in clinical practice, such as imipenem, meropenem and ertapenem, as well as penicillins and cephalosporins, etc. Conclusion: The prevalence of CRE strains in children is increasing year by year, and their antimicrobial resistance to common antibacterial agents in clinical practice is extremely serious, to which serious attention needs to be paid. According to the results of antimicrobial susceptibility testings, the antibacterial agents should be rationally selected to effectively control the spread of CRE.

Elevated expression of CX3C chemokine receptor 1 mediates recruitment of T cells into bone marrow of patients with acquired aplastic anaemia.

OBJECTIVE: Acquired aplastic anaemia (AA) is a T-cell-mediated, organ-specific autoimmune disease characterized by haematopoietic stem cell destruction in the bone marrow. The exact molecular mechanism of T-cell trafficking into the bone marrow is unclear in AA. Very late activation antigen-4 (VLA-4) and CX3C chemokine receptor 1 (CX3CR1) play active roles in many autoimmune diseases. Therefore, we investigated whether VLA-4 and CX3CR1 also contribute to T-cell migration into the bone marrow in acquired AA. DESIGN, SETTING AND SUBJECTS: Expression levels of CX3CR1 and VLA-4 and their ligands [fractalkine (CX3CL1) and vascular cell adhesion molecule-1 (VCAM-1)] were examined in 63 patients with AA and 21 healthy control subjects. T-cell chemotaxis and adhesion were analysed in 17 patients with severe AA. We also prospectively evaluated the expression pattern of CX3CR1 during treatment with antithymocyte globulin plus cyclosporine in 11 patients with severe AA. RESULTS: The proportion of peripheral and bone marrow CD4(+) and CD8(+) T cells expressing CX3CR1 and the level of CX3CL1 was increased in patients with AA. However, there was no significant difference in VLA-4 expression or VCAM-1 levels. Functional studies demonstrated that chemotaxis towards autologous bone marrow plasma or soluble CX3CL1 was significantly higher in T cells from AA patients and could be blocked by CX3CR1 inhibitors. CX3CR1-mediated T-cell adhesion was also upregulated in these patients. The expression of CX3CR1 was associated with the efficacy of immunosuppressive therapy. CONCLUSION: The present findings demonstrate that CX3CR1 plays a pivotal role in recruitment of T cells into the bone marrow in acquired AA and is a potential therapeutic target for treatment of this disorder.

Hypoxia-ischaemia is involved in the pathogenesis of vulvar lichen sclerosus.

BACKGROUND: Lichen sclerosus (LS) is a chronic inflammatory skin disease, the pathogenesis of which is poorly understood. AIM: To evaluate the role of hypoxia-ischaemia (HI) in vulvar LS. METHODS: Samples from five patients with vulvar LS and five control subjects were collected for analysis by transmission electron microscopy (TEM) to reveal the ultrastructural changes of organelles and dermal blood capillaries. Samples from 37 patients with vulvar LS and 12 control subjects were collected for immunohistochemistry to detect the expression of vascular endothelial growth factor (VEGF) and the hypoxia markers hypoxia-inducible factor (HIF)-1alpha and glucose transporter (Glut)-1. RESULTS: Using TEM, the mitochondria of basal cells and vascular endothelial cells in vulvar LS tissue were found to be swollen with loss of cristae, and the rough endoplasmic reticulum had luminal swelling and ribosomal detachment. Damage to vascular endothelial cells, disorganization of capillary architecture and loss of capillaries were also seen. By immunohistochemistry, moderate to intense staining of VEGF was seen in almost 90% of control sections vs. about 55% of LS sections. Glut-1 expression was negative or weak in 75% of control sections vs. moderate to very strong in about 80% of vulvar LS sections. Nuclear staining of HIF-1alpha was not found in LS or control tissue. CONCLUSIONS: HI is involved in the pathogenesis of vulvar LS.

Suppression of glucosylceramide synthase by RNA interference reverses multidrug resistance in human breast cancer cells.

Glucosylceramide synthase (GCS), the enzyme that converts ceramide to glucosylceramide, induce multidrug resistance (MDR) in cancer cells. Recently, RNA interference (RNAi) is a powerful strategy for gene therapy by introducing double-stranded RNA and leading to the sequence-specific destruction. We have designed two different short hairpin RNAs (shRNAs) targeting GCS and introduced them into adriamycin- resistant human breast cancer cells (MCF-7/AdrR cells) to inhibit GCS expression. The results demonstrated that the shRNAs targeting GCS decreased GCS mRNA, abolished GCS protein levels and restored the sensitivity of MCF-7/AdrR cells to several antineoplastic drugs. This study revealed that this approach can reverse MDR effectively and it may be applicable to cancer patients as a specific means to restore the sensitivity to chemotherapy.