The emergence of drug resistance mutations in human immunodeficiency virus (HIV) genome is the major reason for antiviral treatments failure.To investigate the drug-resistance viral quasispecies in HIV-infected individuals and to find out the new drug resistance mutations have significant effects on directing effective clinical treatment and developing novel antiviral drugs.HIV drug resistance tests mainly include genotypic drug susceptibility assay and phenotypic drug susceptibility assay.We reviewed in this article some novel development in the field.

Objective:To investigate the expression and function of CD226 on NK s ubsets and its coexpression with other activation receptor and inhibition recept or on NK cells. Methods:The expression of CD226 on CD56 bright and CD56 dim NK subsets and coe xpress ion with CD16 and NKG2A in PBMC and MLC in the presence or absence of IL-2 or IL-15 were detected by double fluorescent staining and flow cytometry analysis. The level of IFN-? in the supernatants of PBMC culture and MLC treated with o r without IL-2 or IL-15 were evaluated by ELISA. 51Cr release assay w as employed to measure the specific lysis of NK cells killing target K562 cells. Results:CD226 was mainly expressed on CD56 dim NK subsets in PBMC. When s imulated by IL-2, CD226 expression was shifted to CD56 bright NK subsets, while IL-15 increased the subpopulation of NKG2A+CD226+double positive cell s. In MLC-generated NK cells, CD226 was mainly expressed on CD56 dim NK su bsets, and also shifted to CD56 bright NK subsets in the addition of IL-15 . Furt hermore, the percentage of CD16+CD226+和NKG2A+CD226+ subsets were increa sed when stimulated by IL-2 or IL-15. There was great increase in IFN-? lev el in the supernatants of PBMC culture in the presence of IL-2 or IL-15, but no difference in the supernatants of MLC treated with or without two cytokine s. Moreover, the cytotoxicity of NK cells in PBMC and MLC were greatly enhanced by IL-2 or IL-15. Conclusion:CD226 is mainly expressed on CD56 bright NK subsets in IL-2 or IL-1 5 activated NK cells, and is coexpressed with CD16 and NKG2A preferentiatly, whi ch maybe involve in the modulation of cytotoxicity of NK cells based on the balance of coexpressed activation and inhibition receptors. [

Objective:To investigate the expression and function of CD226 on NK cell clone Methods:NK cell clone was established by limited dilution, and identified by FCM The function of CD226 on the cytotoxicity of NK cell clone was detected by RCA and the NK cell clone secreted cytokines in the supernatants during the killing phase were measured by ELISA Results:One NK cell clone was obtained by limited dilution The cytotoxicity of this clone was upregulated markedly by CD226 mAb, and the secretion levels of IFN ? and GM CSF by NK cell clone increased obviously in RCA Conclusion:CD226 is a novel NK cell activation receptor, and the elevated IFN ? and GM CSF levels may be related to CD226 mAb enhanced NK function

Objective Through the analysis of the pathogens distribution of out-patient department and emergency department patients with diarrhea from Shenzhen Hospital of Peking University in 2013~2014,so as to provide reference for the clinical diagnosis and treatment of diarrhea disease.While understanding the improved Salmonella detection results.Methods Var-ied pathogenic bacteria were isolated and identified from 1 719 diarhea stool samples of native Shenzhen Hospital of Peking University from 2013 to 2014 through enrich culture,separate culture,biochemistry,serology etc.Pathogenic virus were test-ed for 451 watery stool specimens by fluorescence PCR.Analyzed statistical differences between the direct inoculation and selenite cystine broth enrichment for Salmonella.Results Picked out 143 disease germs from 1 719 examples diarrhea pa-tient’s stool samples,among which there were 25 strains of ETEC,12 strains of EPEC,8 strains of EAEC,1 strains of EIEC,19 strains ofVibrio parahemolyticus,76 strains Salmonella,2 strains Shigella and 0 strain ofVibrio cholera.There were 10 samples with two disease germs timely.Picked out 189 disease viruses from 451 examples diarrhea patient’s stool without disease germs,among which there were 79 Rotaviruspositive,91 Norwalkvirus positive,9 Adenoviruspositive,10 Astrovirus positive.There were 4 samples with Rotavirus and Norwalkvirus timely.After Salmonella ways to improve the positive rate of 0.6% (17/2 627)increased to 4.4% (76/1 719),χ2=67.2,P<0.01,the difference was statistically signifi-cant.Conclusion The detectable rate of Salmonella and Norwalk virus was the majority,and the clinic enhance the test of the diarrhea pathogenic microorganism,including the improvement of detection method,to reduce the missing rate of them,to provide the scientific basis for the diagnostical therapectic measures.

Objective To investigate the clinical features of cutaneous adverse reactions to tyrosine kinase inhibitors.Methods Thirty patients with cutaneous adverse reactions to tyrosine kinase inhibitors were enrolled from the First Affiliated Hospital of Guangzhou Medical University between January 2015 and December 2016,and their laboratory test results,histopathological findings and treatment response data were collected and analyzed retrospectively.Results Of the 30 patients,15 presented with acneiform eruptions,10 with eczematoid eruptions,2 with morbilliform rashes,1 with telangiectasia,1 with hand-foot skin reaction,9 with xerosis,7 with nail changes and 4 with hair changes.A patient with grade 4 acneiform eruptions showed a markedly elevated alanine transaminase (ALT) level (315 U/L).Mild ALT abnormalities (48.5-88.1 U/L) were found in 3 patients with grade 3 acneiform eruptions,1 with grade 2 acneiform eruptions,1 with grade 1 acneiform eruptions and 1 with eczematoid eruptions complicated by fever.Two patients with eczematoid eruptions and 1 with morbilliform rashes showed elevated proportions of peripheral blood eosinophils (0.057-0.303).Pathological changes of the acneiform eruptions included hyperkeratosis and dilation of hair follicles and neutrophilic infiltration.Pathological manifestations of eczematoid eruptions included different degrees of spongiosis,thickened spinous layer,irregular elongation of rete ridges and liquefaction degeneration of basal cells in the epidermis,and perivascular infiltration of lymphocytes and eosinophils in the superficial dermis.Patients with grade 1-3 acneiform eruptions received oral minocycline for 6 weeks,skin lesions gradually regressed,but relapse occurred after the withdrawal.After withdrawal of targeted antineoplastic agents and 2-week treatment with systemic glucocorticoids,skin lesions gradually regressed in patients with grade 4 acneiform eruptions,those with eczematoid eruptions complicated by fever,and those with morbilliform rashes.Skin rashes also resolved in patients with mild morbilliform rashes and those with mild eczematoid eruptions after 2 weeks of treatment with antianaphylactic agents and topical glucocorticoids.Oral antibiotics were effective for the treatment of periungual erythematous swelling or granulomas.Conclusion Tyrosine kinase inhibitor-related cutaneous adverse reactions include a constellation of disorders,and hepatic function can be impaired.

Objective:To investigate the application values of bone marrow morphology, bone marrow immunohistochemistry, flow cytometry, fluorescence in situ hybridization (FISH) and cytogenetic testing in newly diagnosed multiple myeloma.Methods:A total of 280 patients with multiple myeloma who were newly diagnosed in Tianjin KingMed Diagnosis Center from September 2018 to August 2019 were collected. The bone marrow biopsy was carried out according to the routine method, and bone marrow morphology, bone marrow immunohistochemistry, flow cytometry immunophenotyping, FISH and cytogenetic testing were performed. The detection results of each method were compared.Results:In 280 patients, the bone marrow immunohistochemistry results showed that the median ratio of plasma cells was higher than those of bone marrow morphology (20 cases, 0.675 vs. 0.300) and flow cytometry (47 cases, 0.650 vs. 0.147), and the differences were statistically significant ( Z = -3.883, P < 0.01; Z = -5.947, P < 0.01). Flow cytometry results showed that the positive rates of CD38, CD138, κ, λ, CD56 and CD19 were 100.0% (280/280), 100.0% (280/280), 57.5% (161/280), 42.5% (119/280), 62.1% (174/280) and 19.3% (54/280); bone marrow immunohistochemistry results showed that the positive rates of CD38, CD138, κ, λ and CD56 were 98.9% (277/280), 98.2% (275/280), 57.5% (161/280), 42.5% (119/280) and 62.1% (174/280); there was no statistical difference between the two detection methods in the detection coincidence rate of the same detection index (all P > 0.05). Among patients who underwent FISH detection, the detection rate of gene abnormalities was 69.9% (93/133); the detection rate of abnormalities by direct fluorescence in situ hybridization (D-FISH) was 42.9% (57/133); the detection rate of abnormalities by CD138 immunomagnetic sorting myeloma cells (MACS)-FISH was 82.7% (110/133). Among patients who underwent G-band karyotyping, the detection rate of abnormal karyotype was 38.5% (85/221). FSIH, especially MACS-FISH, had a higher detection rate of cytogenetic abnormalities than G-band karyotyping, and the difference was statistically significant ( χ2 = 65.697, P < 0.05). Conclusion:The comprehensive application of bone marrow morphology, bone marrow immunohistochemistry, flow cytometry, FISH (especially MACS-FISH), cytogenetic testing and other detection methods is more helpful for the diagnosis of multiple myeloma, and may be useful for prognostic judgment.