Objective To study the horizontal transfer mechanism of optrA gene-carrying linezolid-resistant En-terococcus faecalis(LREf)among clinical Enterococcus faecalis mediated by plasmids.Methods Non-repeated LREf from a tertiary first-class hospital in Kunming City from August 2022 to August 2023 were collected and iden-tified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry(MALDI-TOF MS).Antimi-crobial susceptibility testing was performed by VITEK 2 Compact,disc diffusion method and microbroth dilution method,optrA gene was detected by polymerase chain reaction(PCR),molecular biology characteristics of LREf was analyzed by whole-genome sequencing(WGS),LREf as the donor strain and clinically isolated Enterococcus faecalis as the recipient strain for conjugation test.Results A total of 17 LREf strains were collected,optrA gene was detected from plasmids of 12 LREf strains.Multiple resistance genes and virulence genes were detected.12 LREf strains were mainly ST16 type(50.0％).Among the 24 transconjugates in the conjugation test,8 were suc-cessfully conjugated,with a conjugation rate of 33.3％.Further analysis revealed that IS1216E insertion sequences presented at upstream and downstream of the optrA gene on the plasmid before and after conjugation of strains,and formed IS1216E-fexA-optrA-erm(A)-IS1216E-like transposon units.Conclusion The optrA gene can be horizon-tally transferred among clinical Enterococcus faecalis by mediating of plasmid of genes carrying both erm(A)and fexA,and the insertion sequence IS1216E plays an important role in its horizontal transfer process.

Objective To study the horizontal transfer mechanism of optrA gene-carrying linezolid-resistant En-terococcus faecalis(LREf)among clinical Enterococcus faecalis mediated by plasmids.Methods Non-repeated LREf from a tertiary first-class hospital in Kunming City from August 2022 to August 2023 were collected and iden-tified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry(MALDI-TOF MS).Antimi-crobial susceptibility testing was performed by VITEK 2 Compact,disc diffusion method and microbroth dilution method,optrA gene was detected by polymerase chain reaction(PCR),molecular biology characteristics of LREf was analyzed by whole-genome sequencing(WGS),LREf as the donor strain and clinically isolated Enterococcus faecalis as the recipient strain for conjugation test.Results A total of 17 LREf strains were collected,optrA gene was detected from plasmids of 12 LREf strains.Multiple resistance genes and virulence genes were detected.12 LREf strains were mainly ST16 type(50.0％).Among the 24 transconjugates in the conjugation test,8 were suc-cessfully conjugated,with a conjugation rate of 33.3％.Further analysis revealed that IS1216E insertion sequences presented at upstream and downstream of the optrA gene on the plasmid before and after conjugation of strains,and formed IS1216E-fexA-optrA-erm(A)-IS1216E-like transposon units.Conclusion The optrA gene can be horizon-tally transferred among clinical Enterococcus faecalis by mediating of plasmid of genes carrying both erm(A)and fexA,and the insertion sequence IS1216E plays an important role in its horizontal transfer process.

Objective To analyze the influencing factors for intestinal colonization and secondary infection of car-bapenem-resistant Klebsiella pneumoniae(CRKP)in neonates,and provide a basis for formulating prevention and control strategies for CRKP infection.Methods Neonates who were admitted to the neonatal ward of a hospital from January 2021 to October 2022 were selected as the study subjects,and the first screening of CRKP was con-ducted within 48 hours after admission.In addition,active anal swab screening for carbapenem-resistant Ente-robacterales(CRE)was performed weekly during hospitalization,and the infection status of CRKP strains was mo-nitored.Clinical data of neonates in the colonization group,non-colonization group,and infection group were ana-lyzed.Intestinal colonized strains and the non-repetitive CRKP strains isolated from clinical specimens of neonates with secondary infection after colonization were performed carbapenemase gene detection,multilocus sequence ty-ping(MLST)and pulsed-field gel electrophoresis(PFGE)analysis.Results A total of 1 438 neonates were active-ly screened for CRE,174 were CRKP positive,CRKP colonization rate was 12.1％.Among 174 neonates,35 were with secondary infection,with the incidence of 20.1％.The independent risk factors for neonatal CRKP intestinal colonization were cesarean section(OR=2.050,95％CI:1.200-3.504,P=0.009),use of cephalosporins(OR=1.889,95％CI:1.086-3.288,P=0.024),nasogastric tube feeding(OR=2.317,95％CI:1.155-4.647,P=0.018).Protective factors were breast-feeding(OR=0.506,95％CI:0.284-0.901,P=0.021),oral probiotics(OR=0.307,95％CI:0.147-0.643,P=0.002),and enema(OR=0.334,95％CI:0.171-0.656,P=0.001).Independent risk factors for secondary infection after intestinal colonization of neonatal CRKP were carbapenem anti-biotic use(OR=19.869,95％CI:1.778-222.029,P=0.015)and prolonged hospital stay(OR=1.118,95％CI:1.082-1.157,P＜0.001).The detection results of drug resistance genes showed that carbapenemase-producing genes of CRKP strains were all blaKPC-2,all belonged to type ST11.Homologous analysis showed that intestinal CRKP colonization was highly homologous with the secondary infection strains after colonization.Conclusion CRKP intestinal colonization during neonatal hospitalization may increase the risk of CRKP infection.Risk and pro-tective factors of neonatal intestinal colonization and secondary infections after colonization should be paid attention,and corresponding preventive and control measures should be taken,so as to reduce the occurrence and transmission CRKP healthcare-associated infection.

【Objective】 To explore the relationship between the storage time of leukodepleted red blood cells and transfusion adverse reactions by analyzing the occurrence of transfusion adverse reactions of patients after leukodepleted red blood cells transfusion from four hospitals. 【Methods】 By using the electronic medical record management system, the collection and transfusion dates of leukodepleted red blood cells from four hospitals in Enshi Prefecture from 2018 to 2022, as well as the information on transfusion adverse reactions, were retrieved. 【Results】 From 2018 to 2022, a total of 697 61 bags of leukodepleted red blood cells were transfused in four hospitals, resulting in 166 cases of transfusion adverse reactions, among which 93 were allergic reactions, 63 were non hemolytic febrile reactions, and 10 were others, with a total incidence rate of transfusion adverse reactions at 0.24%. The average storage time of leukodepleted red blood cells with and without transfusion adverse reactions was (20.25±6.31) and (19.88±5.50) days, respectively. With a storage time of 7 days as the threshold, the incidence of transfusion adverse reactions was the lowest for a storage time of 15~21 days. The incidence of transfusion adverse reactions of leukodepleted red blood cells in two groups (with storage days ≤21 days and >21 days) was not statistically significant(P>0.05). 【Conclusion】 Allergic reactions were the main type of transfusion adverse reaction caused by leukodepleted red blood cells, and the incidence of transfusion adverse reactions decreased and then increased with the prolongation of the storage time of leukodepleted red blood cells. There was no significant difference in the incidence of transfusion adverse reactions with leukodepleted red blood cells stored for ≤ 21 days and >21 days.

Child ; Humans ; COVID-19 ; SARS-CoV-2 ; Hospitals, Pediatric

Objective:To investigate the characteristics of gene mutation by using whole exome sequencing (WES) and its relationship with therapeutic efficacy in patients with acute myeloid leukemia (AML).Methods:The data of 30 patients with AML from the Second Hospital of Anhui Medical University between December 2014 and September 2019 were retrospectively analyzed; the result of WES, disease type and classification, genetic prognostic stratification and therapeutic efficacy were summarized. The mutation types and mutation frequency of AML patients stratified by the different clinical characteristics and genetic prognosis were compared.Results:Among 30 AML patients, 26 cases (86.7%) had 1 gene mutation at least. The genes with a mutation frequency more than 10.0% were NRAS, RUNX1, TET2, CEBPA, IDH2 and ASXL1. The function of the mutated genes was involved in signaling pathways, transcription factors, epigenetics, and RNA splicing and other biological functions; in terms of mutation pattern, 19 cases (63.3%) of all AML patients mainly presented combined mutations of many combinations. There were no significant differences in mutation rates among age, gender, disease type, disease classification and genetic prognosis stratification groups (all P > 0.05). Of the 8 fusion gene positive cases, 7 were mutated, the average mutation frequency was 175.0% (14/8). There were 19 mutations in 22 patients with negative fusion gene, the average mutation frequency was 213.6% (47/22), and the difference in mutation frequency between the two groups was statistically significant ( P = 0.001). Of the 14 cases of continuous remission, 11 had mutations, with average mutation frequency of 157.1% (22/14); there were 9 cases of mutations in 10 relapsed patients, and the average mutation frequency was 200.0% (20/10); mutations occurred in 5 patients with primary drug resistance, and the average mutation frequency was 300.0% (15/5); and the difference of mutation frequency among the three groups was statistically significant ( P = 0.009). Conclusions:Through WES analysis, it is found that most AML patients have complex and variable gene mutations; and the higher the gene mutation frequency is, the worse the prognosis of the disease is.

OBJECTIVE: To investigate the effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) on the clinical efficacy and flow cytometry (FCM) minimal residual disease (MRD) of patients with acute myeloid leukemia (AML) after initial induction therapy in the real world. METHODS: The clinical data of 44 AML patients who were diagnosed for the first time in the Department of Hematology, The Second Hospital of Anhui Medical University, and received the initial induction therapy were retrospectively analyzed. According to whether rhG-CSF was used after treatment, these patients were divided into control group and therapy group. The complete remission (CR) rate, duration of neutropenia, incidence of infection, duration of fever, cost of antibiotics drugs, length of hospital stay, FCM MRD, and relapse-free survival (RFS) time were compared between the two groups. RESULTS: The CR rate in the control group was 60%, and 74% in the therapy group (P=0.3429). The duration of neutropenia was (21.28±7.91) days in the control group and (14.79±3.07) days in the therapy group (P=0.0016). The duration of fever was (12.80±7.31) days in the control group and (9.11±7.48) days in the therapy group (P=0.0136). While, there were no statistically significant differences between the two groups in the incidence of infection, cost of antibacterial drugs, length of hospital stay and RFS time (all P>0.05). In addition, it is particularly noteworthy that among the patients who finally obtained CR in the therapy group, 66% of them had myeloid precursor cells detected by peripheral blood FCM (accounting for 2.25%±0.99%) at the time of the first release of neutropenia, which was easy to be misdiagnosed as MRD positive. CONCLUSION rhG-CSF not only don't affect the clinical remission rate after the initial induction treatment of AML, but also significantly shortens the time of duration of neutropenia and fever, however, it may affect the analysis of peripheral blood FCM MRD detection results when the neutropenia is released for the first time.
Flow Cytometry ; Granulocyte Colony-Stimulating Factor/therapeutic use* ; Humans ; Induction Chemotherapy/adverse effects* ; Leukemia, Myeloid, Acute/therapy* ; Neoplasm, Residual/etiology* ; Neutropenia ; Recombinant Proteins/therapeutic use* ; Retrospective Studies ; Treatment Outcome

Objective:To seed for stable time window of the integrated disease-syndrome animal model based on the counterevidence from Chinese medicinal prescriptions， and to verify syndrome stability and reliability. Method:A model of depression was established by exposing rats to chronic unpredictable mild stress （CUMS）， followed by body weight measurement， sugar water test， behavioral test， and brain 5-hydroxytryptamine（5-HT） detection. The identification of liver depression and spleen deficiency syndrome was conducted after the equivalent transformation of human clinical symptoms into macroscopic representations of rats. Based on the dynamically collected macroscopic representation scale， Xiaoyaosan was used to reversely verify the stability and reliability of the integrated disease-syndrome animal model of depression due to liver depression and spleen deficiency. Result:The sugar water consumption and the number of crossings and the total movement distance in the open field test of 16-week-old rats in the CUMS （eight weeks of CUMS） group were significantly lower than those in the normal group （P<0.05）. According to the immunohistochemical results， the 5-HT content in hippocampal area CA2 of rats in the CUMS group was also significantly lowered as compared with that in the normal group（P<0.05），which indicated that depression was successfully modeled. The liver depression and spleen deficiency syndrome was present in 14-week-old rats （six weeks after CUMS）of the CUMS group， and the number of rats experiencing the liver depression and spleen deficiency syndrome reached the peak in the 16th week （eight weeks after CUMS），accounting for 70% of the total number. Thereafter， the number decreased gradually. The syndrome scores of the 14-， 16-， 18-， 20-， and 22-week-old rats in the Xiaoyaosan group were reduced by 66.6%， 70.7%， 54.8%， 50.4%， and 44.8%， which were graded as effective， marked effective， effective， effective， and effective， respectively. Conclusion:The age of 14-16 weeks（six to eight weeks after CUMS） is considered the stable and reliable time window for depression due to liver depression and spleen deficiency.

Objective:To study the effect of Wenjing Huayu Zhitong therapy （Xiangyan Zhitong prescription， XZP） on the mitogen activated protein kinase （MAPK）/extracelluar regulated protein kinase （ERK） signaling pathway of primary dysmenorrhea （PD） rats， and explore the pathogenesis of PD and the mechanism of action of Wenjing Huayu Zhitong therapy. Method:Forty-eight female SPF-grade Wistar rats were randomly divided into blank group，model group， western medicine group， low-dose XZP group， medium-dose XZP group， and high-dose XZP group， with 8 rats in each group. In addition to the blank group， dysmenorrhea rat model with cold coagulation and blood stasis syndrome was established by cold stimulation combined with estradiol benzoate and oxytocin. The rats in the blank group，model group，western medicine group， low-dose XZP group， medium-dose XZP group， and high-dose XZP group were given distilled water， distilled water，0.06 g·kg^-1 ibuprofen， 6.55 g·kg^-1 XZP， 13.09 g·kg^-1 XZP， and 26.18 g·kg^-1 XZP， respectively， by gavage for 6 days. The writhing latency and writhing frequency of rats were recorded within 30 min after oxytocin injection.Western blot was used to detect the protein expression of B-Raf， mitogen activates extracellular regulated kinases1/2 （MEK1/2）， extracellular regulated kinases1/2 （ERK1/2）， p-MEK1/2， p-ERK1/2， c-Jun， and cyclooxygenase-2 （COX-2） in rat uterus. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was employed to detect the mRNA expression of B-Raf， MEK1， MEK2， ERK1， ERK2， c-Jun， and cyclooxygenase-2 （COX-2） in rat uterus. Result:Compared with the model group，the treatment groups showed insignificantly prolonged writhing latency and significantly reduced writhing frequency （P<0.01）. On the 6^th day of modeling， there was no significant difference in the quantitative scores of symptoms and signs among the treatment groups. On the 12^th day of modeling， the scores changed little in the western medicine group and the low-dose XZP group and decreased significantly in the medium- and high-dose XZP groups （P<0.01） compared with those in the model group. Compared with those in the blank group， the protein and mRNA levels of p-MEK1/2， p-ERK， B-Raf， c-Jun， and COX-2 in the model group were significantly up-regulated （P<0.01）. Compared with those in the model group， the protein and mRNA levels of p-MEK1/2， p-ERK1/2， B-Raf， c-Jun， and COX-2 in the western medicine group， medium-dose XZP group， and high-dose XZP group were significantly down-regulated （P<0.05，P<0.01）. Conclusion:The mechanism of Wenjing Huayu Zhitong therapy in treating PD with cold coagulation and blood stasis syndrome may be related to the down-regulation of MAPK/ERK signaling pathway.

Kidney cancer usually requires multidisciplinary individualized treatments. No matter what kind of treatment, drugs are essential. According to the "six-step process" (prescription legitimacy review, patient basic information evaluation review, treatment protocol review, organ function and laboratory index review, pretreatment review, and unconventional prescription review) in prescription review proposed by the anti-tumor drug prescription review expert group and referring to domestic and foreign kidney cancer guidelines and drug instructions in recent years, this consensus selects 9 targeted drugs and 4 immunotherapeutic drugs that are currently commonly used in China and elaborates the key review points in patient basic information evaluation review, treatment protocol review, and organ function and laboratory index review of kidney cancer drug treatment, in order to provide reference for clinical front-line pharmacists to review prescriptions of kidney cancer patients and promote rational drug use in clinic.