B cell receptor(BCR)is the main molecular basis for B cell specific recognition and binding of antigen,and its complementarity determining region(CDR3)presents high diversity and specificity.Using single-cell immune repertoire sequencing technology can accurately analyze the composition and characteristics of each B cell BCR CDR3 sequence,and can better understand the process and mechanism of B cell differentiation,development,and response.Single-cell immune repertoire sequencing technology provides new ideas and theoretical basis for the pathogenesis,monitoring,diagnosis and treatment of B-cell immune-related diseases.This paper mainly compares and analyzes the commonly used detection methods and detection platforms of B-cell BCR CDR3 receptor library,and focuses on the application,research status and progress of single-cell sequencing technology in B-cell BCR CDR3 recep-tor library.

Objective To investigate the effects of artesunate(Art)on the proliferation,apoptosis,and autophagy of nephroblastoma cell line(SK-NEP-1).Methods SK-NEP-1 cells were intervened with different concentrations of Art(0,10,20,40 and 80 μmol/L),and MTT method was applied to calculate the cell proliferation inhibition rate to screen the optimal intervention concentration;SK-NEP-1 cells were separated into control group,Art group,3-MA group(Art+autophagy inhibitor,3-methyladenine),and Rapa group(Art+autophagy activator rapamycin).EdU and flow cytometry were applied to detect cell proliferation and apoptosis,respectively;MDC staining was applied to detect autophagy in cells;the level of reactive oxygen species(ROS)in cells was detected by DCFH-DA fluorescent probe;the expression of proliferating cell nuclear antigen(PCNA),anti apoptotic factor B cell lym-phomatoma-2(Bcl-2),Bcl-2 associated X protein(Bax),microtubule junction protein 1 light chain 3 Ⅱ/3 Ⅰ(LC3 Ⅱ/LC3 Ⅰ),selective autophagy junction protein 1(p62),and benzyl chloride 1(Beclin-1)proteins in cells were detected by Western blot.Results Compared with 0 μmol/L Art,the proliferation inhibition rate of SK-NEP-1 cells was gradually increased after 10,20,40 and 80 μmol/L Art treatment(P＜0.05),and the IC50 value was 46.881 μmol/L,so 40 μmol/L Art was selected for follow-up experiments.Compared with the control group,the apoptosis rate,relative autophagy fluorescence intensity,ROS level,Bax,LC3 Ⅱ/LC3 Ⅰ,Beclin-1,PINK1,and Parkin protein expression levels of SK-NEP-1 cells in the Art group were obviously increased,the EdU positive cell rate,PCNA,Bcl-2,and P62 protein expression levels were obviously reduced(P＜0.05);The auto-phagy inhibitor 3-MA inhibited the promoting effect of Art on apoptosis and autophagy of nephroblastoma cells and inhibit proliferation(P＜0.05).Conclusions Art inhibits the proliferation of nephroblastoma cell line SK-NEP-1,and promotes autophagy and apoptosis.

B cell receptor(BCR)is the main molecular basis for B cell specific recognition and binding of antigen,and its complementarity determining region(CDR3)presents high diversity and specificity.Using single-cell immune repertoire sequencing technology can accurately analyze the composition and characteristics of each B cell BCR CDR3 sequence,and can better understand the process and mechanism of B cell differentiation,development,and response.Single-cell immune repertoire sequencing technology provides new ideas and theoretical basis for the pathogenesis,monitoring,diagnosis and treatment of B-cell immune-related diseases.This paper mainly compares and analyzes the commonly used detection methods and detection platforms of B-cell BCR CDR3 receptor library,and focuses on the application,research status and progress of single-cell sequencing technology in B-cell BCR CDR3 recep-tor library.

Objective: Comparing the benefit of Abidor, lopinavir/ritonavir and recombinant interferon α-2b triple combination antiviral therapy and lopinavir/ritonavir and interferon dual combination antiviral therapy to hospitalized novel coronavirus pneumonia 2019 in Zhejiang province. Methods: A multi-center prospective study was carried out to compare the effect of triple combination antiviral therapy with dual combination antiviral therapy in 15 medical institutions of Zhejiang Province. All patients were treated with recombinant interferon α-2b (5 million U, 2 times/d) aerosol inhalation. 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir / ritonavir (2 tablets, 1 time/12 h) as the triple combination antiviral treatment group. 41 patients were treated with lopinavir / ritonavir (2 tablets, 1 time/12 h) as the dual combination antiviral treatment group. The patients who received triple combination antiviral therapy were divided into three groups: within 48 hours, 3-5 days and > 5 days after the symptom onset. To explore the therapeutic effects of triple combination antiviral drugs and dual combination antiviral drugs, as well as triple combination antiviral drugs with different antiviral initiate time. SPSS17.0 software was used to analyze the data. Results: The time of virus nucleic acid turning negative was (12.2 ± 4.7) days in the triple combination antiviral drug group, which was shorter than that in the dual combination antiviral drug group [(15.0 ± 5.0) days] (t = 6.159, P < 0.01 ). The length of hospital stay [12 (9, 17) d] in the triple combination antiviral drug group was also shorter than that in the dual combination antiviral drug group [15 (10, 18) d] (H = 2.073, P < 0.05). Comparing the antiviral treatment which was started within 48 hours, 3-5 days and > 5 days after the symptom onset of triple combination antiviral drug group, the time from the symptom onset to the negative of viral shedding was 13 (10,16.8), 17 (13,22) and 21 (18-24) days respectively (Z = 32.983, P < 0.01), and the time from antiviral therapy to the negative of viral shedding was (11.8±3.9) , (13.5±5.1) and (11.2±4.3) d. The differences among the three groups were statistically significant (Z=32.983 and 6.722, P<0.01 or<0.05). Conclusions The triple combination antiviral therapy of Abidor, Lopinavir/Litonavir and recombinant interferon α-2b showed shorter viral shedding time and hospitalization time compared with the dual combination antiviral therapy. The earlier the time to initiate triple antiviral treatment, the shorter the time of virus shedding.

Objective:To compare the efficacy of the combination of abidol, lopinavir/ritonavir plus recombinant interferon α-2b (rIFNα-2b) and the combination of lopinavir/ritonavir plus rIFNα-2b for patients with COVID-19 in Zhejiang province.Methods:A multicenter prospective study was carried out to compare the efficacy of triple combination antiviral therapy and dual combination antiviral therapy in 15 medical institutions of Zhejiang province during January 22 to February 16, 2020. All patients were treated with rIFNα-2b (5 million U, 2 times/d) aerosol inhalation, in addition 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir/ritonavir (2 tablets, 1 time/12 h) (triple combination group) and 41 patients were treated with lopinavir/ritonavir (2 tablets, 1 time/12 h) (dual combination group). The patients who received triple combination antiviral therapy were further divided into three subgroups: <48 h, 3-5 d and >5 d according the time from the symptom onset to medication starting. The therapeutic efficacy was compared between triple combination group and dual combination group, and compared among 3 subgroups of patients receiving triple combination antiviral therapy. SPSS 17.0 software was used to analyze the data.Results:The virus nucleic acid-negative conversion time in respiratory tract specimens was (12.2±4.7) d in the triple combination group, which was shorter than that in the dual combination group [(15.0±5.0) d] ( t=6.159, P<0.01). The length of hospital stay in the triple combination group [12.0 (9.0, 17.0) d] was also shorter than that in the dual combination group [15.0 (10.0, 18.0) d] ( H=2.073, P<0.05). Compared with the antiviral treatment which was started within after the symptom onset of in the triple combination group, the time from the symptom onset to the viral negative conversion was 13.0 (10.0, 17.0), 17.0 (13.0, 22.0) and 21.0 (18.0, 24.0) d in subgroups of 48 h, 3-5 d and >5 d, respectively ( Z=32.983, P<0.01), while the time from antiviral therapy to viral negative conversion was (11.8±3.9), (13.5±5.1) and (11.2±4.3) d, respectively( Z=6.722, P<0.05). Conclusions:The triple combination antiviral therapy of abidol, lopinavir/litonavir and rIFNα-2b shows shorter viral shedding time and shorter hospitalization time, compared with the dual combination antiviral therapy; and the earlier starting triple combination antiviral therapy will result in better antiviral efficacy.

Objective: To investigate the clinical characteristics and prognostic factors in elderly patients with acute myeloid leukemia(AML). Methods: Clinical data of 232 patients with acute myelocytic leukemia(AML, except for acute promyelocytic leukemia) admitted in our hospital from January 2012 to December 2015 were retrospectively analyzed.Factors affecting complete remission(CR) were analyzed by using χ² test, and Kaplan-Meier survival analysis was conducted.Univariate and multivariate analysis of prognostic factors were performed by using Log-Rank test and Cox regression model respectively. Results: Of 232 patients, 195 patients received induction chemotherapy, among whom 8 patients died in early phase, efficacy could not be evaluated in 25 cases, with 162 patients for final statistical study.The CR rate was 37.0%(60/162) after the first therapy course, and overall CR rate was 54.9%(89/162). Thirty-seven patients received palliative treatment, among whom 6 patients died in early phase and none achieved CR.Therefore, the 162 patients receiving an induction chemotherapy, whose efficacy can be evaluated, could be clinically analyzed.They were in 60-69 years old(χ²=4.102, P=0.043), with ECOG score≤2(χ²=9.917, P=0.002), NPM1⁺ FLT3-ITD^-(χ²=6.423, P=0.038), favorable karyotypes(χ²=6.033, P=0.049), and related to a higher CR rate.The median overall survival(OS) was 205 days in the 232 patients.Univariate analysis results demonstrated that age(χ²=8.700, P=0.003), white blood cell(WBC) count≥100×10⁹/L(χ²=4.249, P=0.039), karyotypes(χ²=4.807, P=0.028), palliative treatment(χ²=191.221, P=0.000) were influencing factors for the prognosis.Multivariable analysis showed that age(HR=0.464, 95%CI: 0.245-0.877, P=0.018), karyotypes(HR=3.618, 95%CI: 1.491-6.728, P=0.003) and whether or not to receive induction chemotherapy(HR=0.076, 95%CI: 0.030-0.194, P=0.000) were independent influencing factors for OS in elderly patients with AML. Conclusions The prognosis of elderly patients with AML is affected by multiple factors.Age, karyotypes and whether or not to receive induction chemotherapy are independent influencing factors for OS in elderly patients with AML.

Accelerated forgetting has been identified as a feature of Alzheimer's disease (AD), but the therapeutic efficacy of the manipulation of biological mechanisms of forgetting has not been assessed in AD animal models. Ras-related C3 botulinum toxin substrate 1 (Rac1), a small GTPase, has been shown to regulate active forgetting in Drosophila and mice. Here, we showed that Rac1 activity is aberrantly elevated in the hippocampal tissues of AD patients and AD animal models. Moreover, amyloid-beta 42 could induce Rac1 activation in cultured cells. The elevation of Rac1 activity not only accelerated 6-hour spatial memory decay in 3-month-old APP/PS1 mice, but also significantly contributed to severe memory loss in aged APP/PS1 mice. A similar age-dependent Rac1 activity-based memory loss was also observed in an AD fly model. Moreover, inhibition of Rac1 activity could ameliorate cognitive defects and synaptic plasticity in AD animal models. Finally, two novel compounds, identified through behavioral screening of a randomly selected pool of brain permeable small molecules for their positive effect in rescuing memory loss in both fly and mouse models, were found to be capable of inhibiting Rac1 activity. Thus, multiple lines of evidence corroborate in supporting the idea that inhibition of Rac1 activity is effective for treating AD-related memory loss.

Objective To investigate the efficacy and safety of domestic bortezomibˉbased chemotherapy for patients with multiple myeloma (MM). Methods The clinical data of 60 MM patients treated with domestic bortezomibˉbased chemotherapy regimen (the observation group) in the First Affiliated Hospital of Zhengzhou University from April 2018 to October 2018 were retrospectively analyzed, which were compared with 112 MM patients treated with original treatment regimen (the control group) at the same hospital from November 2010 to November 2014. According to the disease stage, the patients were divided into newly diagnosed MM (NDMM) group and relapsed refractory MM (RRMM) group, and efficacy and adverse reactions of domestic bortezomib were evaluated. Results The total response rate (ORR) of the observation group was 71.7% (43/60), severe complete response (sCR) + complete response (CR) rate was 16.7% (10/60), very good partial response (VGPR) rate was 18.3% (11/60), and partial response (PR) rate was 36.7% (22/60). The ORR of NDMM group (45 cases) and RRMM group (15 cases) was 82.2% (37/45) and 40.0% (6/15), respectively, and the difference was statistically significant (χ2＝ 9.877, P < 0.05). There was no significant difference between ISS stage Ⅰ+Ⅱ and stage Ⅲ [ORR: 75.7% (28/37) vs. 65.2% (15/23), respectively; χ2＝0.764, P >0.05]. ORR and CR rates in the NDMM group and RRMM group of the observation group and the control group were not statistically different (all P>0.05). In the treatment of bortezomibˉbased chemotherapy, the common adverse reaction was peripheral neuropathy, mostly belonging to grade 1-2. Other side effects included hematocytopenia, gastrointestinal events and herpes zoster, which could be alleviated or restored to normality after supportive treatments. One patient died of pulmonary infection, respiratory failure and septic shock during the intermittent period of chemotherapy. Conclusion ORR of domestic bortezomibˉbased chemotherapy in treatment of the patients with MM is high, and the incidence of adverse reactions shows no significant increase compared with original drugs.

Objective To study the development of the first county-level key clinical specialized talent teams in Henan province. Methods Data on the first 30 county-level key clinical specialized talent teams were collected through questionnaire in 2014 and 2017 respectively, and signed rank sum test was conducted to analyze such facts as the changes of medical service volume, academic titles and scientific researches of discipline leaders before and after such construction. McNemar′s test and chi-square test were conducted to study the age structure, seniority, education and technical title makeup of these discipline leaders. Such tests were also conducted to study the age structure, seniority, education and technical title makeup of the talents. These efforts aim to discern the construction achievement of the key clinical specialties. Results The indexes of healthcare capacities of the first batch of clinical key specialties were observed as improved ( P <0. 05 ) in terms of the medical service volume. Of the discipline leaders′ age structure, the 51 -60 age groups accounted for 40. 0％ and 66. 7％ respectively, before and after the construction, scoring a difference of statistical significance ( P <0. 05 ); the number of senior physicians increased by 80 persons, the number of physicians having master or above titles increased by 40 persons;the number of physicians participating in continuing education held by these specialties grew from 146. 50 to 262. 50(P<0. 05). Conclusions The volume of medical service of the first batch of county-level clinical key specialties has been elevated, and a group of talents of high education and senior titles have been recruited. Whereas, the title structure needs to be improved, and more young academic leaders are expected, while more opportunities of further education and training are also expected for the physicians, in order to enhance their professional ability.

Objective To explore the formation process of vacuum pressure differential of micro power negative pressure technology to facilitate its clinical application.MethodsThe formation of negative pressure differential and changes of negative pressure values in enclosure space were studied which was formed by liquid aspiration after the compressive polyvinyl alcohol foam material was enclosed.Micro power negative pressure technology was applied to the treatment of the right ear necrosis patient after focal cleaning,and the efficacy was observed.Results Medical adhesive film was raised gradually during the expansion of polyvinyl alcohol foam due to liquid aspiration,and then the enclosure space extended increasingly to form local vacuum negative pressure.The range of maximal negative pressure was between-110 and-200 Pa,and the mean maximal negative pressure value was-132.7 Pa.Micro power negative pressure technology behaved well in healing the wound.Conclusion Micro power negative pressure technology gains advantages in reliability and easy operation over other technologies and products,and may be a new choice for healing of refractory wounds.