Objective:To investigate the impact of daily step count on glycemic outcomes in community residents with impaired glucose tolerance (IGT).Methods:This was a prospective cohort study, in October 2018, 204 residents who met the criteria of IGT were recruited in the Shijingshan District in Beijing. The subjects were tested for fasting blood glucose, oral glucose tolerance test 2-hour blood glucose (2hBG), glycated hemoglobin A 1c (HbA 1c), lipid profile, liver and kidney function, as well as measurements of height, weight and waist circumference. A dedicated mobile application was used to deliver prediabetes health knowledge monthly. Online guidance was provided to answer questions and daily step count was collected using the application. Three years later, a follow-up was conducted to assess the participants′ glycemic outcomes and other indexes, and a total of 142 participants completed the follow-up review. According to daily step count, the subjects were categorized into high step count group (42 cases,>7 000 steps daily), moderate step count group (54 cases, 5 000-7 000 steps daily), and low step count group (46 cases,<5 000 steps daily). Subjects were categorized into diabetes group (30 cases), prediabetes group (77 cases) and normal glucose tolerance group (35 cases) with glycemic outcomes. Independent sample t test was used to compare the differences in blood glucose, blood lipids, and step counts between the two groups. Kruskal-Wallis H test or one-way ANOVA was used to compare the differences in blood glucose, blood lipids, and step counts between multiple groups. The χ2 test was used to compare the differences in glycemic outcomes between multiple groups. Multivariate logistic regression analysis was used to assess the impact of daily step counts and body mass index on glycemic outcomes. Linear regression analysis was used to evaluate the relationship between daily step counts and 2 h BG. Results:A total of 142 participants completed the 3-year follow-up, including 43 males and 99 females, with a mean age of (60.15±5.67) years. At baseline, males had significantly higher body mass index, waist circumference, and fasting blood glucose when compared to those in females [(26.97±2.43) vs (24.89±2.93) kg/m 2, (92.68±7.75) vs (83.83±8.60) cm, (5.83±0.61) vs (5.62±0.52) mmol/L], the total cholesterol and HDL-C were also significantly lower in males than those in females [(5.10±1.16) vs (5.55±0.95) mmol/L, (1.35±0.34) vs (1.56±0.35) mmol/L] (all P<0.05). After 3-year follow-up, 21.1% (30/142) of IGT participants progressed to diabetes, with an annual conversion rate of approximately 7%. The normal glucose tolerance group showed significantly higher daily step counts when compared with the prediabetes and diabetes groups [(7 886±2 867) vs (5 981±2 655) vs (4 117±2 674) steps] ( H=31.778, P<0.001). Individuals with higher daily step counts exhibited lower body mass index, 2 h BG, and HbA 1c level when compared with those in the ones with moderate and low step counts [(24.26±3.09) vs (25.44±3.38) vs (26.26±3.59) kg/m 2, (7.50±1.71) vs (9.15±3.30) vs (11.19±3.84) mmol/L, 5.97%±0.46% vs 6.14%±0.99% vs 6.40%±0.96%] (all P<0.05). Higher step count was positively correlated with the reversal of prediabetes to normal blood glucose levels (moderate step count, OR=0.297, 95% CI: 0.109-0.804; low step count, OR=0.055, 95% CI: 0.010-0.287), lower daily step count correlated positively with prediabetes progressing to diabetes ( OR=4.857, 95% CI: 1.140-20.689) (all P<0.05). For every additional 1 000 steps per day, the 2 h BG decreased by 0.5 mmol/L. Conclusion:As daily step count increases, the glucose metabolism improves in IGT community residents. Higher daily step count is associated with reversal of IGT to normal glucose tolerance, while lower daily step count may be associated with the progression of IGT to diabetes.

Objective:To construct and prepare recombinant virus strains chimeric with human metapneumovirus (HMPV) antigenic epitopes.Methods:Recombinant influenza virus vectors which chimeric with different HMPV antigenic epitopes were rescued by reverse genetics using eight-plasmid system. The recombinant influenza virus strain used the internal genes of A/PR/8/34 (PB1, PB2, PA, NP, NS, M, HA, and NA) as a backbone, with concomitant genetic modifications to insert the B-cell epitopes of HMPV into the HA gene, and the CTL+ Th cell epitopes of HMPV into the NA gene. Preparation of recombinant influenza virus strains using reverse genetics in a " 7+ 1" model. The recombinant virus strains were evaluated by measuring hemagglutinin (HA) titers, half tissue culture infectious dose (TCID 50) and growth curves. Sequencing analysis was conducted to verify whether the rescued viruses carried the chimeric HMPV epitopes. Results:The epitopes of HMPV were inserted into the influenza virus genome and two recombinant influenza virus strains were rescued successfully, named as FLU/HMPV/B and FLU/HMPV/CTL+ Th. HA titers of the recombinant strains were both 2 7, their TCID 50 were 10 5.2/ml and 10 5.0/ml, respectively. After cultured for three passages in chick embryo, these two recombinant strains could proliferate steadily. Whole genome sequencing verified that the FLU/HMPV/B carried the B-cell epitopes of HMPV, the FLU/HMPV/CTL+ Th carried the CTL and Th cell epitopes of HMPV. Growth curve tests also verified that the recombinant strains could proliferate steadily in eggs. Conclusions:Two recombinant influenza virus vector strains carrying the B cell, CTL and Th epitopes of HMPV were rescued successfully. The result of the recombinant virus strains in terms of growth characteristics as well as genetic stability indicate that they meet the requirements for proceeding to the next step of animal experiments. The immunogenicity and immunoprotective effect will be further evaluated by mouse experiments. Ultimately new ideas for the realization of " one vaccine for two uses" or " one vaccine formultiple uses", as well as a new strategy for the development of HMPV vaccine will be proposed.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective:To understand the epidemiological characteristics of group A rotavirus (RVA) infection in hospitalized children under 5 years of age with diarrhea in 2019, and to provide reference for the surveillance of RVA.Methods:Stool samples and clinical information of hospitalized children under 5 years of age with diarrhea were collected from sentinel hospitals in 20 provinces in 2019. RVA nucleic acid detection and genotyping were performed according to the rotavirus detection method in the National Viral Diarrhea Surveillance Program.Results:A total of 5 395 viral diarrhea samples were collected, 5 038 were tested, and 1 247 diarrhea samples showed RVA positive results (1 247/5 038, 24.75%). The positive rate of RVA in Fujian province was the lowest (30/319, 9.40%), and the positive rate of RVA was the highest in Henan province (182/338, 53.85%). The positive rate of RVA in male and female children was 25.24%(762/3 019)and 24.02%(485/ 2 019), respectively. There was no significant gender distribution of RVA infection ( χ2 = 0.96, P=0.326). Children aged 12 to 17 months were mainly susceptible to RVA (342/1 033, 33.11%), and the positive rate of RVA in children aged 48 to 59 months was lower (35/227, 15.42%). RVA infection showed significant age distribution characteristics ( χ2 = 86.78, P<0.001). RVA infection had significant difference between urban and rural areas ( χ2 = 20.92, P<0.001) and seasonal characteristics ( χ2 =411.42, P<0.001). RVA genotyping showed that G9P[8] type (994/1 122, 88.59%) was the dominant epidemic strain. Conclusions:In 2019, the main genotype of RVA infection in hospitalized children under 5 years of age with diarrhea was G9P[8], and RVA infection had significant age, region and season characteristics.

Objective:To design the fourth-generation chimeric antigen receptor-T (CAR-T) cells that secrete interleukin-7 (IL7) and chemokine C legend 19 (CCL19) on the basis of the second-generation CAR, and to analyze and compare the differences in proliferation, chemotaxis, tumor cell clearance and persistence in the microenvironment of multiple myeloma (MM) between them.Methods:The fourth-generation CAR vector plasmid was constructed by using 2A self-cleaving peptide technology. The third-generation lentiviral packaging system was used to prepare high-titer lentivirus. Flow cytometry was used to monitor the transduction efficiency of lentivirus and the subtype changes of CAR-T cells. The enzyme-linked immunosorbent assay (ELISA) was used to quantify the IL7 and CCL19 secreted by CAR-T cells.The calculation of absolute number of CAR-T cells during culture was used to analysis cell proliferation activity. Transwell migration assay was used to verify the chemotactic ability of CAR-T cells. The specific killing activity of CAR-T cells was detected by using the luciferase bioluminescence method. The NOD-Prkdcem26Cd52Il2rgem26Cd22/Nju (NOD) mouse xenograft model was used to verify the anti-myeloma activity and safety of CAR-T cells in vivo.Results:Flow cytometry results showed that the stable CAR expression rates of the second-generation anti-CS1 CAR-T and fourth-generation anti-CS1-IL7-CCL19 CAR-T cells were (91.50±0.29)% and (46.7±0.12)%, respectively. CAR-T cells were successfully constructed. Subtype analysis demonstrated that the ratio of stem memory T cell (TSCM) in anti-CS1-IL7-CCL19 CAR-T cells was (67.58±0.59)%, which was significantly higher than (50.74 ± 1.01)% of anti-CS1 CAR-T ( P=0.000 1), with more strong immune memory function and better durability. Anti-CS1-IL7-CCL19 CAR-T cells can continuously secrete IL7 and CCL19 compared to MOCK-T and anti-CS1 CAR-T ( P<0.000 1). The number of anti-CS1-IL7-CCL19 CAR-T cells reached (22.77±0.79)×10 6 on the 9th day after lentivirus transduction, which was significantly higher than (9.40±0.79)×10 6 of anti-CS1 CAR-T cells ( P=0.000 1), with stronger proliferation ability. The number of chemotaxis cells of anti-CS1-IL7-CCL19 CAR-T cells to reactive T cells was (109.0±4.04), which was significantly higher than (9.33±1.20) of MOCK-T ( P<0.000 1) and (7.33±0.88) of anti-CS1 CAR-T ( P<0.000 1), with stronger chemotactic ability. The specific killing activity showed that both anti-CS1-IL7-CCL19 CAR-T and anti-CS1 CAR-T cells had specific killing efficacies when compared with the MOCK-T cells ( P<0.000 1). Animal experiment indicated that anti-CS1-IL7-CCL19 CAR-T cells significantly reduced the tumor burden ( P<0.000 1) and extended the overall survival time ( P=0.006 1) of tumor-bearing mice. Conclusions:The anti-CS1-IL7-CCL19 CAR-T cells designed in this study show stronger proliferative activity, chemotactic ability, and durability without affecting the anti-myeloma activity in vivo and in vivo, which provides strategies for overcoming the defects of low survival rate, poor durability and inhibition by tumor microenvironment of traditional CAR-T cells, and offers preliminary experimental basis for the clinical application of the fourth-generation CAR-T cells.

Objective:To design the fourth-generation chimeric antigen receptor-T (CAR-T) cells that secrete interleukin-7 (IL7) and chemokine C legend 19 (CCL19) on the basis of the second-generation CAR, and to analyze and compare the differences in proliferation, chemotaxis, tumor cell clearance and persistence in the microenvironment of multiple myeloma (MM) between them.Methods:The fourth-generation CAR vector plasmid was constructed by using 2A self-cleaving peptide technology. The third-generation lentiviral packaging system was used to prepare high-titer lentivirus. Flow cytometry was used to monitor the transduction efficiency of lentivirus and the subtype changes of CAR-T cells. The enzyme-linked immunosorbent assay (ELISA) was used to quantify the IL7 and CCL19 secreted by CAR-T cells.The calculation of absolute number of CAR-T cells during culture was used to analysis cell proliferation activity. Transwell migration assay was used to verify the chemotactic ability of CAR-T cells. The specific killing activity of CAR-T cells was detected by using the luciferase bioluminescence method. The NOD-Prkdcem26Cd52Il2rgem26Cd22/Nju (NOD) mouse xenograft model was used to verify the anti-myeloma activity and safety of CAR-T cells in vivo.Results:Flow cytometry results showed that the stable CAR expression rates of the second-generation anti-CS1 CAR-T and fourth-generation anti-CS1-IL7-CCL19 CAR-T cells were (91.50±0.29)% and (46.7±0.12)%, respectively. CAR-T cells were successfully constructed. Subtype analysis demonstrated that the ratio of stem memory T cell (TSCM) in anti-CS1-IL7-CCL19 CAR-T cells was (67.58±0.59)%, which was significantly higher than (50.74 ± 1.01)% of anti-CS1 CAR-T ( P=0.000 1), with more strong immune memory function and better durability. Anti-CS1-IL7-CCL19 CAR-T cells can continuously secrete IL7 and CCL19 compared to MOCK-T and anti-CS1 CAR-T ( P<0.000 1). The number of anti-CS1-IL7-CCL19 CAR-T cells reached (22.77±0.79)×10 6 on the 9th day after lentivirus transduction, which was significantly higher than (9.40±0.79)×10 6 of anti-CS1 CAR-T cells ( P=0.000 1), with stronger proliferation ability. The number of chemotaxis cells of anti-CS1-IL7-CCL19 CAR-T cells to reactive T cells was (109.0±4.04), which was significantly higher than (9.33±1.20) of MOCK-T ( P<0.000 1) and (7.33±0.88) of anti-CS1 CAR-T ( P<0.000 1), with stronger chemotactic ability. The specific killing activity showed that both anti-CS1-IL7-CCL19 CAR-T and anti-CS1 CAR-T cells had specific killing efficacies when compared with the MOCK-T cells ( P<0.000 1). Animal experiment indicated that anti-CS1-IL7-CCL19 CAR-T cells significantly reduced the tumor burden ( P<0.000 1) and extended the overall survival time ( P=0.006 1) of tumor-bearing mice. Conclusions:The anti-CS1-IL7-CCL19 CAR-T cells designed in this study show stronger proliferative activity, chemotactic ability, and durability without affecting the anti-myeloma activity in vivo and in vivo, which provides strategies for overcoming the defects of low survival rate, poor durability and inhibition by tumor microenvironment of traditional CAR-T cells, and offers preliminary experimental basis for the clinical application of the fourth-generation CAR-T cells.

Objective To analyze special histopathological characteristics of melanocytic nevi and their associations with age,gender,anatomical locations and pathological subtypes.Methods Clinical and histopathological data were collected from 1 011 patients with melanocytic nevi,who visited Beijing Hospital from January 2005 to January 2019,and analyzed retrospectively.Statistical analysis was carried out by using chi-square test for comparing enumeration data,and t test for comparing measurement data.Results Among the 1 011 patients with melanocytic nevi,the age at the clinic visit was 40.90 + 19.19years,and there were 289 males and 722 females.Lesional (biopsy) sites included the trunk (402 cases,39.8％),face and neck (268 cases,26.5％),extremities (138 cases,13.6％),hands and feet (133 cases,13.2％),scalp (53 cases,5.2％) and vulva (17 cases,1.7％).Pathological subtypes included intradermal nevus (580 cases,57.4％),compound nevus (333 cases,32.9％) and junctional nevus (98 cases,9.7％).Among special histopathological characteristics,neuralization and adipose cell hyperplasia were observed in 172 (17.0％) and 155 (15.3％) cases respectively,and the prevalence of neuralization and adipose cell hyperplasia was significantly higher in female patients than in male patients,higher in elderly patients than in young patients,and higher on the scalp than on the other sites (all P ＜ 0.05);vascular proliferation was observed in 313 (31.0％) cases,and more commonly occurred on the scalp than on the other sites (P＜0.05);nevus cells distributed along the hair follicles/sebaceous glands were observed in 502 (49.7％)cases,and more commonly seen on the face and neck than on the other sites (P ＜ 0.05);nevus cell lysis occurred in 203 (20.1％) cases,and fissures were observed in 384 (38.0％).All the above histopathological characteristics were more frequently observed in the intradermal nevus subtype than in the compound nevus subtype (all P＜0.05).Nevus cells distributed along the blood vessels were observed in 20 (2.0％) cases,and more commonly seen on the extremities than on the trunk,hands and feet (P ＜ 0.05),as well as in the compound nevus subtype than in the intradermal nevus subtype (P ＜ 0.05).Conclusions There are many special histopathological characteristics in melanocytic nevi,such as neuralization,adipose cell hyperplasia,vascular proliferation,and nevus cells distributed along the hair follicles/sebaceous glands,which are associated with patients' age,gender,lesional locations and histopathological subtypes.

Objective To investigate clinical features of vulvar lichen simplex chronicus (VLSC).Methods Clinical data were collected from 137 VLSC patients who visited a Vulvar Clinic in the Department of Dermatology,Beijing Hospital from 2017 to 2018,and analyzed retrospectively.Non-normally distributed measurement data (age and disease duration) were described as median (P25,P75),and analyzed by using rank sum test,and enumeration data were compared by using chi-square test.Results Among the 137 patients with VLSC,the age at onset was 32.0 (25.5,40.0) years,and the disease duration was 36.0 (15.0,72.0) months.Thirty-two (23.4％) patients had a history of atopic diseases,and the age at onset of VLSC was significantly lower in these patients with a history of atopic diseases (29.5 [25.0,35.8]years) than in those without a history of atopic diseases (33.0 [27.0,41.0] years,Z =2.03,P =0.042).The most frequently involved site was the labia majora (130/137,94.9％),and the labia minora was rarely involved (13/137,9.5％).Bilateral lesions were observed in 103 (75.2％) patients,and hypopigmentation occurred in 8 (5.8％) patients.All the patients experienced itching to different extents,including moderate itching in 44 (32.1％) cases and severe itching in 80 (58.4％).The ratio of patients with severe itching to those with disease duration ＞ 2 years (68.1％) was significantly higher than that of patients with severe itching to those with disease duration ＜ 2 years (47.7％,x2 =5.830,P =0.016).Patients reported that local wetness and sweating (55 cases,40.1％),spicy diet (41 cases,29.9％) and mental stress (36 cases,26.3％) could aggravate itching,Conclusions VLSC commonly occurs in patients aged 20-39 years,and atopic predisposition may be an important factor for VLSC.VLSC mostly involves bilateral labia majora,and the longer the disease duration,the more severe the itching.

Objective To analyze correlations of the distribution of non-segmental vitiligo lesions with gender,age of onset,duration of disease,personal and family history of autoimmune diseases,family history of vitiligo,and so on.Methods Clinical data were collected from 1 125 patients with confirmed nonsegmental vitiligo in Department of Dermatology,Beijing Hospital from January 2009 to January 2019,and analyzed retrospectively.Statistical analysis was carried out with SPSS 20.0 software by using independentsample t test and chi-square test.Results Of the 1 125 patients with non-segmental vitiligo,599 were males and 526 were females,with a male-to-female ratio of 1.14:1.Their age of onset was 27.9 ± 17.1 years,and duration of disease was 5.2 ± 8.0 years.Skin lesions mostly occurred on the trunk (544 cases,48.4％) and face (535 cases,47.6％),followed by acral regions (430 cases,38.2％),extremities (297 cases,26.4％) and neck (231 cases,20.5％).The perioral region (17.2％),hands (47.9％) and genital region (14.5％) in the 599 male patients were more frequently affected than those in the 526 females (9.7％,22.6％,6.3％,x2 =13.33,77.66,20.01,respectively,all P ＜ 0.001),while the neck (27.0％) was more frequently affected in the female patients than in the male patients (14.9％,x2 =25.29,P ＜ 0.001).The age of onset was significantly lower in the patients with vitiligo lesions involving the legs,knees,feet,trunk,genital and periocular regions than in those without skin lesions on the above corresponding body sites (all P ＜ 0.05),but significantly higher in the patients with vitiligo lesions involving the arms and hands than in those without lesions on the above 2 sites (both P ＜ 0.05).Vitiligo lesions more likely occurred on the hands of patients with a personal or family history of autoimmune thyroid diseases compared with those without the personal or family history (50.0％ vs.27.9％,x2 =6.62,P =0.010).The trunk was more frequently affected in the patients with a family history of vitiligo than in those without (59.6％ vs.45.7％,x2 =13.36,P ＜ 0.001).Conclusion The distribution of non-segmental vitiligo lesions are correlated to some extent with gender,age of onset,personal and family history of autoimmune diseases and family history of vitiligo.

Objective:To analyze changes in melanocyte density and epidermal thickness in vulvar lichen sclerosus (VLS) lesions.Methods:Vulvar skin tissues were collected from 15 adult female patients with VLS in Department of Dermatology, Beijing Hospital from June to December in 2018. According to pathological manifestations, 15 skin lesions were divided into early-stage VLS group ( n = 7) and late-stage VLS group ( n = 8) , and subjected to immunofluorescent staining. Then, density of melanocytes, and thickness of the whole epidermis and cell layers (from the bottom of the stratum corneum to the bottom of the basal layer) were calculated by using an image analysis software. Normal vulvar skin tissues were obtained from 15 healthy adult females after vulvar plastic surgery in Plastic Surgery Hospital, Chinese Academy of Medical Sciences, and served as control group. Results:The density of melanocytes in the epidermis was significantly lower in the early-stage VLS group (0.170 ± 0.071) and late-stage VLS group (0.110 ± 0.035) than in the control group (0.275 ± 0.036; F = 36.426, P < 0.001) . There was no significant difference in the thickness of the whole epidermis among the early-stage VLS group (203.682 ± 137.997 μm) , late-stage VLS group (150.020 ± 70.914 μm) and control group (194.030 ± 82.996 μm; F = 0.738, P = 0.487) . The thickness of cell layers did not differ between the early-stage VLS group (154.603 ± 121.984 μm) and control group (176.974 ± 80.296 μm; P = 0.899) , but was significantly thinner in the late-stage VLS group (83.455 ± 37.129 μm) than in the control group ( P = 0.003) . Conclusions:The density of epidermal melanocytes decreased in the early-stage and late-stage VLS skin lesions. Compared with the normal skin tissues, the early-stage VLS lesions showed no significant changes in the thickness of the whole epidermis and cell layers, but the late-stage VLS lesions showed decreased thickness of cell layers.