Objective To assess the predictive value of neutrophil gelatinase associated protein lipocalin (uNGAL) in urine for detection of acute kidney injury(AKI) in patients with severe traumatic brain injury. Methods Patients with severe traumatic brain injury from the ICU were collected from Jan. 2011 to May. 2013 in our hospital. 43 cases that met the RIFLE criteria for diagnosis of AKI in the ICU within 7 days were selected as AKI group. Another 43 cases that were matched for age ,gender ,illness severity , surgery method with AKI cases ,selected as non‐AKI group. The levels of uNGAL and Scr were measured when the patients admit‐ted in the ICU with 15 min ,at 24 h ,48 h ,72 h. the sensitivity and specificity of uNGAL and Scr for diagnosis for AKI were evalua‐ted by ROC curve. Results The incidence of severe traumatic brain injury AKI was 42. 16% (43/102). The uNGAL levels in the AKI group were higher when the patient stayed in the ICU longer and no obvious in the non AKI group. When admitted to the ICU 24 h ,the level of uNGAL(720. 32 ± 684. 25)ng/mL in AKI group was significantly higher than that (421. 92 ± 351. 20)ng/mL in non AKI group. The difference was statistically significant (P< 0. 05). The levels of Scr between two groups were not statistically significant. The area under ROC curve of uNGAL and Scr were 0. 879 (95% CI :0. 807 - 0. 949) and 0. 612 (95% CI :0. 493 -0. 731). When the cutoff value of uNGAL was 180 ng/mL ,the sensitivity and specificity were 0. 890 and 0. 823 respectively. The sensitivity was superior to Scr. Conclusion uNGALis superior to Scr for early diagnosis of AKI in patients with severe traumatic brain injury and it could be used as a biomarker for early diagnosis of AKI.

Objective To study the value of flow cytometry in identifying metastatic CK positive and negative nonhematopoietic neoplasms in bone marrow. Methods Twenty-six cell lines representing ten epithelial neoplasms, one lymphoma cell line and one human T cell lymphoblast-like cell line were purchased from American Tissue Culture Collection. From July 2009 to June 2010, five nonhematopoietic neoplasms,fifteen hematopoietic neoplasms and fifteen control patients with complete remession after hematopoietic stem cell transplantation were collected in Beijing Daopei Hospital. Cryopreserved cell lines were thawed and cultured until they entered log phase. After permeabilization, cell lines were analyzed by staining with cytoplasmic CK-FITC antibody using four-color flow cytometer. The percent CK positivity was measured by comparing with negative control. Bone marrow samples were stained with membrane and cytoplasmic antibodies according to our routine methods. Based on lineage markers and blast markers as well as CK expression, the relevant hematopoietic diseases were diagnosed or excluded according to 2008 World Health Organization diagnosis standards. Results All epithelial neoplasm cell lines expressed CK, with average positive percentage 81.1%. All the lymphoid tumor cell lines didn't express CK. Two epithelial neoplasms were CK positive, 100. 0% in thyroid carcinoma and 98. 2% in lung carcinoma, respectively. Hematopoietic tumor and control samples didn't express CK. They expressed relevant hematopoietic markers, such as CD45 as well as lineage markers, or CD138 and cytoplasmic immunoglobulin light chain. Three nonepithelial nonhematopoietic neoplasms didn't express CK. CK positive or negative nonhematopoietic neoplasms didn't express hematopoietic markers such as CD45, HLA-ABC and HLA-DR DP DQ, as well as lineage specific markers. Besides, CK positive might be helpful to suggest epithelial origin. Conclusion Flow cytometry with hematopoietic markers and CK can effectively exclude hematopoietic tumor and identify metastatic CK positive and negative nonhematopoietic neoplasms in bone marrow.

ObjectiveTo understand the characteristics of mutations in BCR-ABL1 kinase domain mutation,these chronic myeloid leukemia (CML) and Ph positive acute lymphoblastic leukemia (ALL)patients who got imatinib treatment had poor effect.MethodsTotally 177 CML patients and 33 Ph( + )ALL patients were selected at Beijing Dao-Pei Hospital from Sep.2007 to Dec.2010.All of them were Chinese patients.Totally 243 bone marrow or peripheral blood specimens were collected from the patients,who had early effect,then resistance emergenced,or for more than 3 months of poor efficacy.Extracted total RNA from the specimens' nuclear cells,reversed transcription to cDNA.Amplified the whole span of BCRABL1 fusion kinase gene by nest PCR (from 242 to 493 amino acid coding sequence),used the type AB3130XL gene sequencing instrument determinate the gene sequence of ABL1 kinase region and then used the Variant Reporter V1.0 software to analyze the results of gene mutations.ResultsThirty-two kinds of different mutations were detected of ABL1 gene mutations,accounting for 34.2％ (83/243 cases).Among them,the T315I was 12％ (10/83),mutation rate was the highest,followed by Y253H was 11％ (9/83),G250E was 7％ (6/83),E255K was 7％ (6/83),M351T was 6％ (5/83),E459K was 5％ (4/83) ;Q252H,D276G,F317L,E355G,F359V,H396R were all 4％ (3/83).Three cases of insertion mutations were found,including 2 cases of 357-358insk,1 case of V304RfsX17.Seven patients had found existence two or more point mutations.The multiple drug resistance mutations might exist in the same leukemia clone.The same individual was not only contain common resistance mutations,but also rare point mutations,insertion mutations.The mutations might be lead to loss of kinase activity.ConclusionsUnder the imatinib drugs pressure,the ABL1 gene mutation in leukemia cells appears randomly,and results in different resistant clones.Different resistant clones can coexist in the same patients in vivo; resistant clones not only contain point mutations,but also contain inserted deletion mutations.

Objective: To analyze the pathogenic spectrum of infectious diarrhea among children in Zhengzhou City in 2020, so as to provide insights into the management of childhood infectious diarrhea. Methods: A total of 230 stool samples were collected from children with diarrhea in the Diarrhea Clinic of Zhengzhou Municipal Children's Hospital in 2020. Diarrheagenic Escherichia coli, Salmonella, Shigella, Vibrio parahaemolyticus, Campylobacter and Aeromonas were isolated. Salmonella was typed using pulsed-field gel electrophoresis ( PFGE ) and antimicrobial resistance was tested in Salmonella. The main types and drug resistance of pathogens were descriptively analyzed. Results: The 230 children with diarrhea included 152 males and 78 females, with a male to female ratio of 1.95∶1. There were 173 cases at ages of 3 years and below ( 75.22% ), and 82 cases seen between June and August ( 35.65% ). A total of 71 pathogenic bacteria strains were isolated, with a detection rate of 30.87%. Salmonella was the most frequently isolated pathogen ( 35 strains, 15.22% prevalence ), following by diarrheagenic E. coli ( 20 strains, 8.70% ). Eleven serotypes of Salmonella were identified, which mainly included S. enteritidis and S. typhimurium. PFGE typing revealed 25 band patterns of Salmonella, with a similarity ranging from 11.85% to 100%. Salmonella were sensitive to imipenem, meropenem, polymyxin B and amikacin and highly resistant to streptomycin ( 85.71% ), ampicillin ( 80.00% ) and sulfaisoxazole ( 80.00% ). A 82.86% proportion of multi-drug resistance was detected in the 29 Salmonella strains. Conclusions A wide range of pathogenic bacteria were detected from the childhood infectious diarrhea in Zhengzhou City in 2020, which mainly included Salmonella and diarrheagenic E. coli. A high polymorphism level was seen in PFGE fingerprints of Salmonella, which presented high-level resistance to antimicrobial agents.

Objective: To investigate the serotypes and drug resistance of non-typhoidal Salmonella in Zhengzhou City, so as to provide insights into prevention and control of non-typhoidal Salmonella infections. Methods: Salmonella isolates were collected from diarrheal patients in Zhengzhou municipal sentinel hospitals from 2017 to 2021. Salmonella serotypes were identified using slide agglutination test and soft agar colony formation assay, and antimicrobial susceptibility test was performed using the broth micro-dilution method. Results: Five serogroups and 37 serotypes were identified among 446 non-typhoidal Salmonella isolates, with S. enteritidis (210 isolates, 47.09%) and S. typhimurium (133 isolates, 29.82%) as dominant serotypes. Non-typhoidal Salmonella showed high resistance to ampicillin (79.60%), ampicillin/sulbactam (58.74%), naphthyric acid (56.05%), tetracycline (54.26%) and doxycycline (54.04%), respectively. There were 290 multidrug-resistant Salmonella isolates (65.02%), and the multidrug resistance rates were 70.48% for S. enteritidis and 67.67% for S. typhimurium, respectively. Conclusions Multiple serotypes of non-typhoidal Salmonella were identified in Zhengzhou City from 2017 to 2021, with S. enteritidis and S. typhimurium as dominant serotypes. Widespread drug resistance and multidrug resistance was seen in non-typhoidal Salmonella.

Objective To develop a multiplex sequence-specific PCR assay for simultaneous screening of 5 types of JAK2 mutations and investigate its clinical application value. Methods Multiplex sequence-specific PCR assay for simultaneous screening of JAK2 V617F, K539L (include 2 types of gene mutations), N542-E543del and E543-D544del mutations were developed. 115 patients with myeloproliferative diseases (MPD) including 61 polycythemia vera (PV) cases, 43 essential thrombocythemia (ET) cases and 11 primary myelofibrosis (MF) cases were analyzed. Results The assay can screen the 5 types of JAK2 mutations efficiently. The detection sensitivity is 1% for JAK2 V617F mutation and 0.1% for the other mutations. JAK2 V617F mutation and JAK2 exon12 mutation were detected in 56 and 3 of the 61 PV samples, respectively. 27 of the 43 ET samples and 6 of the 11 MF samples were JAK2 V617F positive, but no JAK2 exon12 mutation was found in both groups. The 3 cases carrying JAK2 exon12 mutation had the clinical feature of erythrocytosis and erythropoietin-independent erythroid colony formation but without apparent leukocytosis, thrombocytosis and splenectasis. Conclusion The assay can simultaneously screen 5 types of JAK2 mutations with high sensitivity and thus lead to an increased detection rate.

Objective To analyze the etiological factor and genetic feature of a familial hemophagocytic lymphohistiocytosis patient with PRF1 mutation (FHL2) with human herpesvirus 7 (HHV7)infection and its family constellation. Methods Clinical characteristics, laboratory examinations of a FHL2 case with HHV7 infection were reported. HHV1-HHV8 virus DNA was screened by PCR; NK cell function was analyzed by flow cytometry; PRF1 gene mutations were analyzed by PCR and direct sequencing, structure of mutant PRF1 proteins were analyzed using ExPasy and I-TASSER server and genetics pedigree were analyzed. Results The patient's HHV7 viral was detected positive with DNA copy number of 350/106 peripheral nucleated cells. Flow cytometry analysis showed decrease both in proportion of perforin positive NK cells and perforin protein expression. Genetic testing showed PRF1 biallelic heterozygote mutations (c. 503G ＞ A/p. S168N and c. 1177T ＞ C/p. C393R) and pedigree analysis showed they were inherited. The patient was then treated with antivirus therapy, dexamethasone and VP16 therapy, but only achieved partial response. The patient was then followed by human leukocyte antigen 10/10 allele identical nonconsanguinity allogeneic hematopoietic stem cell transplantations (allo-HSCT) and soon the successful implantation of donor hematopoietic cells and persistent recovery was achieved. The patient was now surviving without recurrence for 9 months after allo-HSCT. Conclusions FHL is prone to be misdiagnosed as lymphoma. Genetic analysis of related gene mutation and herpes simplex virus detection will help in early and accurate diagnosis. Allo-HSCT is a fundamental treatment of FHL.

BACKGROUNDTo investigate whether autologous lung cancer tissues derived gp96-peptide complex/dendritic cell vaccine could induce peptide specific cytotoxic T lymphocyte (CTL) response in vitro.

METHODSA patient's tumor-derived antigens including gp96-peptide complexes and tumor cell lysate were co cultured with DCs derived from the same patient's bone marrow blood mononuclear cells. The various antigen/DC vaccines were used to stimulate peripheral lymphocytes. Interferon-γ (IFN-γ) level of activated lymphocytes was detected by ELISA method and the Cr51 release test was performed to evaluate the gp96-peptide specific CTL response in three kinds of target cells including the primary cultured tumor cells, PG cells and K562 cells.

RESULTSIFN-γ could be observed from the supernate collected in all antigen groups after the cognate T lymphocytes were stimulated by various vaccines. The concentration of IFN-γ induced by gp96-peptide complexes/DC vaccine was higher than that of other groups. In addition, the killing effect of the activated T lymphocytes on patient's primary tumor cells was higher than that on PG and K562 cells.

CONCLUSIONSAutologous tumor-derived gp96-peptide complexes can induce a peptide complex specific CTL response, and the CTL response is significantly intensified after DCs are pulsed.

Objective To investigate the clinical efficacy and prognosis of lung transplantation in the treatment of cystic fibrosis (CF). Methods Clinical data of one patient with end-stage CF undergoing allogeneic bilateral lung transplantation were retrospectively analyzed. Clinical characteristics, diagnostic methods and treatment strategies of the CF recipient were summarized. Results The recipient had suffered from relevant symptoms since childhood including repeated cough and purulent sputum for 30 years, complicated with recurrent pulmonary infection combined with acute exacerbation, chronic sinusitis and extremely severe malnutrition. Prior to lung transplantation, the patient had to depend upon the invasive ventilator due to respiratory muscle weakness, and admitted to intensive care unit (ICU) for a long time. Imaging examination revealed multiple cystic columnar bronchiectasis accompanied with infection in bilateral lungs. The diagnosis of CF was further confirmed by sweat test and gene detection. The recipient underwent bilateral lung transplantation on August 17, 2017 and received rehabilitation treatment. The lung function was gradually restored to normal. The recipient had obtained the same quality of life to the healthy counterparts since the date of manuscript submission (over 2 years). Conclusions Lung transplantation is an efficacious treatment for end-stage CF, which can not only save patients' lives, but also significantly improve the quality of life of patients.

Objective To study the type and corresponding clinical characteristics of primary hemophagocytic lymphohistocytosis (HLH) associated immune gene mutations in the refractory virus infection or HLH of unknown causes. Methods From December 2009 to July 2010, the patients with refractory virus infection or HLH of unknown causes were screened for the primary HLH associated immune genes mutations by DNA sequence analysis, including PRF1, UNC13D, STX11, STXBP2, SH2D1A and XIAP. The clinical characteristics and outcomes were followed up. Results Totally 25 patients with refractory virus infection or HLH of unknown causes were investigated for the 6 genes and 13 cases were found carrying gene mutations, composing of 6 of PRF1 mutation, 3 of UNC13D, and each one of STX11,XIAP, SH2D1A and STXBP2, respectively. Among the 13 cases with gene mutations, 5 suffered from Epstein-Barr virus associated HLH( EBV-HLH), 1 human herpes virus 7 associated HLH (HHV7-HLH),1 HLH without causes, 4 chronic activated EB virus infection (CAEBV) with 1 progressing to Hodgkin's lymphoma carrying abnormal chromosome of t ( 15; 17 ) (q22; q25 ) and hyperdiploid, 2 EBV associated lymphoma. Among the other 12 patients without gene mutation, 4 suffered from EBV-HLH with 1 progressing to peripheral T lymphoma, 8 suffered from CAEBV. Conclusions Primary HLH associated immune gene mutations are critical causes of refractory virus infection of unknown causes, most patients manifest as HLH,some cases appear in CAEBV and EBV associated lymphoma. DNA sequence analysis is helpful to early diagnosis and correct decision-making for treatment.