Objective To investigate the diagnostic value of immunophenotype in distinguishing acute promyelocytic leukemia(APL)from HLA-DR negative acute myeloid leukemia(AML)using flow cytometry.Methods A retrospective observational study was con-ducted including 42 APL patients and 28 newly diagnosed or relapsed HLA-DR negative AML patients admitted to our hospital from 2014 to 2024.Immunophenotype analysis was performed on bone marrow aspirate samples using flow cytometry.The positive expression rates of CD64,MPO,CD7,CD11c,CD9,CD123 and other antigens were compared between the two groups using the Chi-square test.The diagnostic efficiency of the CD9/123 and CD64+MPO+CD7 CD11c-models for APL was evaluated using receiver operating charac-teristic(ROC)curves.Results The HLA-DR negative AML group exhibited significantly lower positive rates of CD64,CD9 and MPO(P＜0.05),and higher positive rates of CD11c and CD7(P＜0.05)compared to APL group.The CD64+MPO+CD7-CD11c-model had an area under the curve(AUCROC)of 0.859,sensitivity of 93.8％and specificity of 75.0％for distinguishing APL.The CD9/CD123 expression pattern had AUCROC of 0.919,sensitivity of 83.3％and specificity of 84.0％for APL diagnosis.The combined CD9/123 and CD64+MPO+CD7-CD11c-model had AUCROC of 0.955,sensitivity of 83.3％and specificity of 100％.Conclusion The combined CD9/123 and CD64+MPO+CD7-CD11c-expression pattern may serve as a helpful tool for differentiating APL from HLA-DR negative AML.

Objective To analyze the interventional effect of high-flow nasal cannula oxygen (HFNC) therapy in patients with stable chronic obstructive pulmonary disease (COPD) combined with hypercapnia. Methods A total of 45 patients with stable COPD complicating with hypercapnia who require long-term oxygen therapy were selected as study subjects. They were divided into three groups based on different respiratory support modes: long-term home oxygen therapy (LTOT) group, non-invasive ventilation (NIV) group, and HFNC group, with 15 patients in each group. The general condition, blood gas index, lung function index, respiratory status, quality of life, and walking test results of the three groups were compared after discharge. Results During the follow-up period, the number of admission and acute exacerbation in the HFNC group and NIV group was lower than that in the LTOT group (P < 0.05). At 6 and 12 months after discharge, the arterial partial pressure of carbon dioxide [p_a(CO₂)] in the three groups was lower than before discharge, while the arterial partial pressure of oxygen[p_a(O₂)], pulse oxygen saturation (SpO₂), forced vital capacity (FVC), and forced expiratory volume in one second (FEV₁) were higher than before discharge. Moreover, the changes in the above indicators in the HFNC group were greater than those in the NIV group and LTOT group (P < 0.05). At 1-, 3-, 6-, and 12-month after discharge, the scores of dyspnea scale of the modified Medical Research Council (mMRC) and the St. George′s Respiratory Questionnaire (SGRQ) in the three groups gradually decreased, and the 6-minute walking distance (6MWD) gradually increased. Additionally, the changesin the above indicators in the HFNC group were greater than those in the NIV group and LTOT group (P < 0.05). Conclusion HFNC can effectively alleviate dyspnea symptoms, improve lung function, promote the recovery of blood gas indicators, enhance quality of life and exercise tolerance, and reduce re-hospitalization rates in patients with stable COPD combined with hypercapnia.

Objective:To analyze the correlation factors between peripapillary duodenal diverticulum (PDD) and choledochectasia by CT scan.Methods:The clinical data of 220 patients with duodenal diverticulum detected by multi-slice spiral CT scan and confirmed by gastrointestinal angiography or endoscopic retrograde cholangiopancreatography (ERCP) in Dahua Hospital, Xuhui District of Shanghai City were retrospectively analyzed. The correlation of the PDD, the contact of common bile duct (CBD), length of contact and exudation with choledochectasia in patients with PDD were analyzed.Results:A total of 236 duodenal diverticulum were found in 220 patients. Among them, there were 152 PDD, 41 diverticulum located superior to the duodenal papilla, 28 diverticulum located inferior to the duodenal papilla, 3 diverticulum located lateral to the duodenal papilla, and 12 diverticulumlocated in the horizontal portion. The incidence of choledochectasia in patients with PDD contacted with CBD was significantly higher than that in patients with PDD not contacted with CBD: 59.35% (73/123) vs. 37.93% (11/29), and there was statistical difference ( P<0.05); the incidence of choledochectasia in patients with contact length of PDD and CBD ≥1.5 cm was significantly higher than that in patients without contact of PDD and CBD and patients with contact length of PDD and CBD <1.5 cm: 82.43% (61/74) vs. 24.49% (12/49) and 37.93% (11/29), and there was statistical difference ( P<0.05); the incidence of choledochectasia in PDD patients with exudation was significantly higher than that in PDD patients without exudation: 10/11 vs. 52.48% (74/141), and there was statistical difference ( P<0.05). Conclusions:The patients with contact length of PDD and CBD ≥1.5 cm and patients with PDD combined with exudation could be prone to choledochectasia.

Objective:To evaluate the skin development and repair process of X-ray radiation damage in rat with non-invasive two-photon excitation fluorescence (TPEF) imaging technology in vivo. Methods:Totally 24 SD rats were randomly divided into four groups including X-ray irradiated group (25, 35 and 45 Gy) and non-irradiation control group. At different times after irradiation, the degree of skin injury was evaluated, and the pathological changes of nicotinamide adenine dinucleotide (phosphate) [NAD(P)H] and collagen fiber fluorescence signals in epidermal cells were detected in vivo by TPEF imaging technology. Results:At 10 d post-irradiation, the skin of irradiation groups showed erythema and desquamation. At 15-20 d post-irradiation, the skin of radiation groups developed progressive exudation, edema and ulcers with increasing radiation dose. On day 25, the skin began to repair in the 25 Gy group, however, the skin of other groups still had exudation and ulcers. On day 10, NAD(P)H fluorescence signal in epidermal cells of irradiation groups decreased and the fluorescence signal of collagen fibers in papillary layer and reticular layer of irradiation groups reduced, which were significantly lower than that of normal control group ( t=24.145, 28.303, 26.989, 6.654, 7.510, 7.997, P<0.05). On day 30, fluorescence signal of NAD(P)H and collagen fibers in epidermal cells and dermis began to repair, the cell from stratum granulosum, stratum spinosum, and stratum basale in the 25 Gy group showed fluorescence signal, the other groups did not show. The fluorescence signal of collagen fibers in the 25 Gy group were gradually increased in papillary layer and reticular layer, however, they were significantly lower than normal control group ( t=115.133, 17.431, P<0.05), the skin of 45 Gy group did not show fluorescence signal of collagen fibers. Conclusions:The damage and repair process of epidermal cells and dermal collagen fiber can be detected noninvasively by TPEF imaging technology after X-ray irradiation in vivo.

Teaching design is an important part of the construction of teaching contents,and it plays an important role in improving the quality of education and teaching. At present, teaching design has not received enough attention in the teaching of the physical chemistry for the Chinese pharmacy major. This study discussed the importance of the teaching design in the classroom of physical chemistry for the Chinese pharmacy major and expounded some practices of teaching design in detail, including the design of the affective attitude goals for the purpose of improving students' sense of curriculum identity and stimulating students' interest in learning, the design of "points-lines-planes" for teaching and learning based on the mind map,and the design of the ways of the classroom lead-in such as case-based learning mode.

In order to analyze the immunogenicity of the recombinant EG95s protein ,the recombinant plasmids of pET-1EG95s ,pET-2EG95s and pET-3EG95s which containing respectively 1 ,2 ,and 3 copies EG95s were induced to express HIS-1EG95s ,HIS-2EG95s and HIS-3EG95s ,and then the proteins were purified and identified by western-blotting .The same im-mune process was used ,and 8 weeks-old BALB/c mice were immunized ,then its immunogenicity was analyzed by detecting an-tibody levels in mice by indirect ELISA method .Results showed that for recombinant EG95s proteins after transformation , HIS-1EG95s ,HIS-2EG95s ,and HIS-3EG95s also retained immunogenicity and could induce specific antibodies in mice .One week's late after the first immunization with HIS-1EG95s ,the antibody level of was significantly higher than HIS-2EG95s and HIS-3EG95s .But began from 2 weeks after immunization ,the antibody level of HIS-3EG95s was always higher than that of HIS-1EG95s group during the period of the immune .Both the final antibody titers after immunization of HIS-1EG95s and HIS-2EG95s groups was 1∶819 200 ,while HIS-3EG95s group was 1∶163 840 0 .HIS-1EG95s ,HIS-2EG95s and HIS-3EG95s all induced IFN-γin immune mice ,but the difference was not significant .The HIS-1EG95s showed lower response to Echinococ-cus granulosus positive serum than HIS-2EG95s and HIS-3EG95s .It’s indicated that the HIS-1EG95s and HIS-3EG95s also had good immunogenicity .HIS-3EG95s make recombinant protein immunic effects more lasting ,and benefit to generate more long-lasting protective immunity .This study provides the scientific basis for the immunization of echinococcosis (hydatidosis) .

Objective To explore the different apoptotic gene expressions and apoptotic signaling transduction of human rhabdomyosarcoma (RD) cells infected by enterovirus 71 (EV71) in different stage.Methods The survival of EV71-infected RD cells was observed by trypan blue staining.The apoptotic morphology and rates of RD cells were surveyed and detected by Annexin V-FITC/PI staining and flow cytometry,respectively.PCR array was employed to analyze 88 apoptotic gene expressions from EV71-infected RD cells at 8 h and 20 h postinfection (p.i),respectively.Results After RD cells were infected with EV71 (MOI =5) at 8 h p.i,the viability was significantly decreased.Flow cytometry data demonstrated that the apoptotic rates of EV71-infected RD cells had increased to 18.0％ and 19.1％ at 20 h p.i in early and later stage respectively.RT-PCR array studies revealed significant variations in the expression of apoptotic genes.Among 88 genes,only the expression of IFN-β1 was upregulated 5.22 folds,whereas 47 genes including ACIN1,Akt,Apaf1,caspase and CIDEB were found to be downregulated that were lower than 2 folds at 8 h p.i.However,28 genes including FasL,CD40L,TNF,caspase-10 and caspase-3 were induced more than 2 folds after EV71 infection at 20 h.Conclusion The downregulation of apoptosis-related genes is associated with viral apoptosis-suppressing effect in RD cells in the early stage of EV71 infection.The death receptor signaling pathways including Fas/FasL and TNF/TNFR are activated to induce cell apoptosis in the late stage of EV71 infection.Moreover,host cell can also inhibit apoptosis by regulating signal pathway of CD40/CD40L,NF-κB/RelA and PI3K/Akt activation.

Objective To detect Chlamydia trachomatis phage Vp1 gene in clinical swab specimens and anti-Vp1 antibodies in serum specimens.MethodsCervical and urethral swab as well as serum specimens were collected from attendees to the sexually transmitted disease(STD) clinic in the Tianjin Institute of STD,during March 2008 to March 2011.PCR was conducted to detect chlamydial phage Vp1 gene in swab samples,enzyme linked immunosorbent assay(ELISA) and Western blot to detect anti-Vp1 antibody in sera.The swab specimens positive for Vp1 gene were subjected to cell culture followed by the detection of Vp1 protein with an immunofluorescence-based method.ResultsTotally,36 out of 1542 swab specimens turned out to be positive for Vp1 gene,and 23 out of 453 serum specimens for anti-Vp1 antibody.No positive results were obtained in the Vp1 gene-positive swab specimens by cell culture and immunofluorescence-based assay.ConclusionThe Vp1 gene of Chlamydial trachomatis phage and anti-Vp1 antibody are successfully detected from clinical swab and serum specimens respectively.

Objective To get phagc's capsid Vp3 gene and protein of guinea pig inclusion conjunctivitis (GPIC) chlamydia. Methods The genome DNA was extracted from the φCPG1 phage.The full sequence of Vp3 gene was amplified by polymerase chain reaction (PCR) from the above genome DNA. The Vp3 gene was digested by restriction endonuclease and then inserted into prokaryotic plasmid vector pET30a (+). The recombinant plasmid was transformed into E. coil BL21, and was identified by restriction endonuclease, PCR and sequencing. The E. coil BL21 with expected recombinant plasmid was induced and the expressed recombinant Vp3 protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, then purified by agarose gel. Results The recombinant gene was sequenced and proved to be 447 bp which was consistent with the φCPG1 Vp3 gene sequence in GenBank. A 25 000 capsid protein was expressed and confirmed by SDS-PAGE and Western blot. The purified protein was obtained. Conclusion The capsid Vp3 protein of φCPG1 is successfully expressed and purified, which is helpful for the further study on its mechanism and clinical applications.

Objective To investigate specific immune responses in mice induced by recombinant major outer membrane protein(rMOMP)of C.trachomatis serovaf E.Methods Thirty-six female BALB/cmice aged 3 to 4 weeks Were divided into three groups.i.e.,adjuvant group vaccinated、with purified rMOMP and Freund's adjutant,solitary group vaccinated with rMOMP only and control group vaccinated with phosphate buffered saline(PBS).All the mice were intramuscularly vaccinated on week 0,2 and 4.Blood samples and vaginal washes were obtained from these mice on week 6,then,mice were challenged with elementary body(EB)of C.trachomatis serovar E at the footpad followed by the observation of delayed hypersensitivity.On week 7.mice were genitally infected with C.trachomatis EB;one week later,blood samples and vaginal washes were obtained again;six weeks later,spleen lymphocytes were isolated from the mice and stimulated bv C.trachomatis or ConA followed by the detection of cell proliferation with MTT assay.In vitro neutralization assay was also performed.ELISA was used to determine the titers of Chlamydia-specific IgO antibody in sera and IgA antibody in vaginal washes,as well as the level of IFN-γ in culture supernatant of lymphocytes and sefa of mice.Vaginal swabs were collected after genital challenge and subjected to C.trachomatis culture.Results The absorbance at 405 ms of Chlamydia-specific IgG antibody and proliferation index of lymphocytes were 0.641±0.059 and 5.085±1.291.respectively,in mice immunized with rMOMP and Frennd's adjuvant.significantly higher than those in mice immunized with rMOMP only(0.424±0.015 and 3.123 ±0.840.both P<0.05).The thickness of right hind footpad increased by 0.324±0.054 mm and 0.272±0.064 mm,respectively,in solitary group and adjuvant group,respectively,with significant difference between the two groups(P<0.05).A significant increase was also observed in the adjuvant group compared with the control group in the above three parameters(all P<0.01).Conclusion The rMOMP of C.trachomatis could efficiently induce Chlamydia-specific humoml and cellular immune responses in mice.