A 11-year old female patient with severe thalassemia, receipt a lentivirus-based cell and gene therapy (CGT) therapy in Shenzhen Children′s Hosptial on July 27th, 2021. At the two follow-up visits after discharge, patient were continuously tested positive for HIV screening through HIV Ag/Ab Combo assay (chemiluminescence Immunoassay), and the viral load results of HIV-1 nucleic acid testing (NAT) were both>5 000 copies/ml. The patient can be diagnosed with HIV infection according to the National Guideline for Detection of HIV/AIDS(2020 Revised Edition). The thorough investigation findings and supplementary experiment results indicated that the false-positive HIV-1 NAT results was caused by cross-reactivity between the target sites detected by conventional HIV-1 NAT reagents and the lentiviral vectors fragments integrated into the genome of patient′s hematopoietic stem/progenitor cells. In conclusion, it is important for laboratories to select appropriate HIV-1 NAT testing platforms which won′t cause cross-reactivity for the testing of samples from patients who have been treated with HIV-derived vectors. It is also recommended to design and develop NAT testing platforms with multiple target regions labeled by different fluorescents for HIV NAT supplementation experiment to reduce the risk of false-positive diagnoses of HIV infection.

Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine

Objective:To evaluate the immunogenicity and protective effect of a multicomponent recombinant protein vaccine EPRHP014 constructed independently and provide a scientific basis for developing new tuberculosis (TB) vaccine and effective prevention and control of TB.Methods:Three full-length Mycobacterium ( M.) tuberculosis protein antigens (EsxH, Rv2628, and HspX) and two epitope-predicted and optimized epitope-dominant protein antigens (nPPE18 and nPstS1) were selected, from which five protein antigens were used to construct a protein antigen composition EPRHP014, including a fusion expression multi-component protein antigen (EPRHP014f) and a multi-component mixed protein antigen (EPRHP014m) formed with the five single protein using clone, purification, and purification respectively. Multicomponent protein vaccines EPRHP014f and EPRHP014m were prepared with aluminum adjuvant, and the BCG vaccine was used as a control. ELISA detected the titer of serum-specific antibodies, the secretion of various cytokines was detected by ELISpot and Luminex, and immune protection was observed by the M.tuberculosis growth inhibition test in vitro. The results were statistically analyzed by t-test or rank sum test, and P<0.05 was considered a statistically significant difference. Results:Mice Immunized with EPRHP014m and EPRHP014f could produce highly effective IgG antibodies and their subtypes IgG1 and IgG2a, and the antibody titers were similar to those of mice immunized with BCG, with no statistical significance ( P>0.05). The number of spot-forming cells (SFC) secreting IFN-γ and IL-4 induced by EPRHP014f group was significantly higher than those by EPRHP014m group and BCG group ( P<0.05), but there was no significant difference in the number of SFC for IFN-γ and IL-4 induced between EPRHP014m group and BCG group ( P>0.05). The secretion levels of GM-CSF and IL-12p70 induced by the EPRHP014m group were higher than those of the BCG group ( P<0.05), but there was no significant difference in the levels of IL-6 and IL-10 induced between EPRHP014m group and BCG group ( P>0.05). There was no significant difference in the secretions of IL-6, IL-10, IL-12, and GM-CSF between the EPRHP014f and BCG groups ( P>0.05). EPRHP014m group, EPRHP014f group, and BCG group had obvious antibacterial effects in vitro, and the difference was insignificant ( P>0.05). Conclusion:Both EPRHP014f and EPRHP014m can induce strong humoral and cellular immune responses in mice after immunization, and have a strong ability to inhibit the growth of M. tuberculosis in vitro, indicating that the antigen composition EPRHP014 has good potential in the development and application of TB vaccine.

Objective:To analyze the resistance mutational profiles of multi-drug resistant Mycobacterium tuberculosis in China and the correlation between major mutation types and genotypes based on the whole-genome sequencing data. Methods:Search and download of the genome-wide sequencing data of M. tuberculosis published in China by August 2019 on NCBI database were conducted. Mutation frequency of drug resistance-related gene loci based on whole-genome sequencing was used to predict the molecular susceptibility of strains, and the correlation between mutation types and genotypes was analyzed. Results:According to the results of molecular resistance and susceptibility profiles, 1 024 MDR strains were identified from 2 019 M. tuberculosis strains. The major mutation types of resistance-related genes to common drugs were katG S315T (73.2%, isoniazid), rpoB S450L (63.1%, rifampicin), rpsL K43R (70.0%, streptomycin), embB M306V (37.4%, ethambutol), pncA_promoter T (-11)C (7.9%, pyrazinamide), gyrA A90V (32.3%, fluoroquinolones), rrs A1401G (67.7%, second-line injection drugs), fabG1_promoter C (-15) T (87.0%, Ethionamide), folC I43T (30.4%, P-aminosalicylic acid). Among them, the frequencies of katG S315T, embB M306V, rpsL K43R, gyrA A90V in lineage 2 were significantly higher than those in lineage 4, and folC I43T was only found in lineage 2. The proportion of katG S315T was significantly higher in the ancient Beijing genotype compared to the modern genotype, in contrast, the proportion of rpsL K43R was significantly higher in modern Beijing genotype, the differences were significant (all P<0.05). Conclusions:The results showed the main mutation types of resistance-related genes of MDR strains to many commonly used anti-tuberculosis drugs in China based on whole-genome sequencing, providing a basis for the development of sensitive and specific rapid molecular detection methods. At the same time, it was also found that the major mutation types of MDR-related genes were related to the genotype of the strains.

Objective:To explore the real experience of family caregiver in providing care service and receiving home care organizational management after the disabled aged enjoyed the long-term care insurance so as to provide a scientific basis for standardizing the long-term care insurance home care service.Methods:From July to September 2019, this study selected 14 family caregivers of the home disabled aged enjoying the long-term care insurance in Guangzhou by purposive sampling to carry out the semi-structured interview so as to understand the personal feelings and suggestions of interviewees on long-term care insurance home care service. Colaizzi phenomenological analysis was used to data analysis.Results:A total of two themes were abstracted including the harvest of family caregiver and insufficient management model.Conclusions:Long-term care insurance home care service is instrumental in reducing the economic burden, care burden of family caregivers, and promoting the normative development of home care service. However, there is insufficient management model of family caregivers which needs to be improved.

Objective:To explore benefit finding in patients receiving combined heart-lung transplantation (CHLT) , and provide a reference for improving clinical nursing work.Methods:From September to October 2019, totally 6 patients who survived after CHLT in the Department of Cardiac Surgery at the First Affiliated Hospital of Guangzhou Medical University were selected by purposive sampling for semi-structured interviews in terms of their benefit finding using phenomenological methods. Colaizzi's seven-step analysis was used to organize and analyze the interview results.Results:The patients undergoing CHLT showed a sense of benefit finding, which could be summarized into four themes, namely, changes in mental quality (learning to be grateful and contented, understanding the meaning of life, learning to be open-minded and optimistic) , improved learning initiative (actively learning drug-related knowledge, the knowledge of rehabilitation and exercise, the knowledge of diet and nutrition, and the knowledge of prevention of complications) , perceiving social support (family members changed life habits, family members actively learned disease knowledge, improved communication between patients and family members and friends) , and actively planning future life.Conclusions:The patients receiving CHLT can experience a sense of benefit finding in the rehabilitation after organ transplantation. Medical staff should understand the sense of benefit finding in patients receiving CHLT, promote the early generation of benefiting finding, and provide targeted guidance to patients.

Objective To study the predictive value of umbilical cord blood regulatory T cells (Treg) for bronchopulmonary dysplasia (BPD) in preterm infants. Method From June 2017 to December 2018, premature infants with gestational age less than 32 weeks admitted to NICU of our hospiatal were prospectively selected. The umbilical cord blood was collected at birth to examine the Treg amount. The infants were assigned into BPD group and non-BPD group according to the diagnosis at discharge. The differences of Treg amount between the two groups and different degrees of BPD were analysed. Result A total of 124 premature infants (GA<32 weeks) were admitted, including 41 cases in BPD group (mild, n=18;moderate, n=14; severe, n=9) and 83 cases in the non-BPD group. The BPD group had GA of (29.6 ± 1.1) weeks and birth weight (BW) of (1128 ± 135) g. The non-BPD group had GA of (29.8 ± 1.1) weeks and BW of (1316 ± 180) g. The birth weight, 1min and 5min Apgar scores in BPD group were lower than the non-BPD group (P<0.001). The BPD group had higher incidence of respiratory distress syndrome, longer duration of mechanical ventilation (MV) and oxygen inhalation(P<0.001) than the non-BPD group. The MV duration and oxygen inhalation duration in the severe BPD group were significantly longer than the moderate and mild BPD groups, and the duration in the moderate group was longer than the mild group (P<0.001). The number of Treg in cord blood in the BPD group [(1.43 ± 0.06) × 105 cells/ml] was significantly lower than the non-BPD group [(2.57 ± 0.09) × 105 cells/ml], and the difference was statistically significant (P<0.001). Multivariate Logistic regression analysis showed that a significant decrease in the number of Treg was a risk factor for BPD in premature infants (OR=0.000, 95％CI 0.000 ～ 0.012, P=0.009). The number of Treg in umbilical cord blood was negatively correlated with the severity of BPD. The area under the ROC curve showed that the cut-off value was 1.95 × 105 cells/ml, with Youden index 0.613, sensitivity 85.4％ and specificity 75.9％. Conclusion The number of cord blood Treg cells may be a useful biomarker for predicting BPD in premature infants.

Objective To investigate the value of F-actin autoantibodies in the serum of patients with systemic lupus erythematosus (SLE),and to explore the relationships between F-actin autoantibodies and other clinical indicators.Methods ELISA was established to detect serum levels of F-actin autoantibodies in 93 inpatients with SLE from March 2017 to January 2018 (case group,n=93),72 patients with rheumatoid arthritis (RA) (disease control group) and 83 healthy subjects (healthy control group) were included during the same period.The positive rates of F-actin autoantibodies between the case group and the two control group were compared.Clinical data including SLE disease activity index (SLEDAI),immuno-globulin (lg)G,erythrocyte sedimentation rate (ESR),anti-dsDNA,and antinuclear antibody (ANA) of 93 patients with SLE were collected and the correlation analysis between F-actin autoantibodies units was applied respectively.The diagnostic performance of F-actin autoantibodies in SLE was analyzed by using the receiver operating characteristic curve (ROC).T test,Chi-square test and Spearman/Pearson correlation analysis were applied for statistical analysis.Results The serum levels of F-actin autoantibodies in the SLE case group,disease control group,and healthy control group were (18±13),(12±6),and (11±5) U,respectively,the differences between SLE case group and disease control group,and healthy control group were significant (t=3.163,P=0.001 9;t=4.436,P＜0.01).The positive rates of F-actin autoantibodies were 33％(31/93) in patients with SLE,10％(7/72) in disease control group,and 4％(3/83) in healthy control group.The F-actin autoanti-bodies units in SLE were correlated with SLEDAI,IgG,ESR,anti-dsDNA,and ANA (r=0.273 7,P=0.008 3;r=0.558 7,P＜0.01;r=0.419 9,P=0.000 1,r=0.351 4,P=0.001 1,r=0.460 9,P＜0.01),in which F-actin autoantibodies units showed significant correlation with IgG and ANA.In the ROC curve,the area under the curve(AUC) was 0.62 [95％CI(0.54,0.70)],P=0.001 3.which was statistically significant.When the cut-off value of the F-actin autoantibodies was 14.04 U,the Youden's index (YI) was the largest (YI=0.30),and the sen-sitivity for the diagnosis of SLE was 0.77,the specificity was 0.53.Conclusion The positive rate of F-actin autoantibodies in the serum of patients with SLE is higher than that of RA and healthy controls,so it has certain diagnostic value for SLE.The F-actin autoantibodies units is correlated with both SLEDAI,ESR,and anti-dsDNA,suggesting that F-actin autoantibodies units may be a new biomarker for disease activity assessment of SLE patients.

Objective To establish an extended nursing service model for intravenous treatment in outpatient,so that patients with venous catheters nearby can receive catheter maintenance,which can reduce the medical cost and the incidence of complications during the maintenance of the catheters.Methods A three-level network of catheter maintenance was established in the area to evaluate the rate of return maintenance and the incidence of complications [catheter-related infection,medical adhesive-related skin injuries (MARSI) and catheter blockage] during catheterization.Results The rate of patient return maintenance as statistically signif-icant between the first half of 2015 and the second half of 2016 (60.0％ vs.29.7％,P=0.000).Compared with the first half of 2015,the incidence of catheter-related infections decreased in the second half of 2016 from 8.6％ to 3.3％,the incidence of MARSI decreased from 29.7％ to 10.1％,and the incidence of catheter blockage decreased from 8.5％ to 0.6％.(All P =0.000).Conclusion The comprehensive extended service mode for outpatient with venous catheters can reduce the incidence of complications during catheter maintenance,which can reduce the workload of outpatient intravenous treatment and also provide patients the convenience of catheter maintenance,and is worthy of clinical promotion.

Objective To investigate the effect of different doses of ioversol on renal function,and to explore early renal injury biomarkers on contrast induced kidney injury and safe ioversol dosage.Methods A total of 158 cases (98 males and 60 females) undergoing cerebral vascular intervention (CVI) in our department was selected with age ranging from 23 to 81 years old (average age 59.70 ± 12.02).Based on ioversol dosage in surgery,patients were divided into three groups:low dose group (≤ 150 ml,n =49),middle dose group (151-200 ml,n =74),and high dose group (＞200 ml,n =35).U-κ,U-λ,urinary transferrin (UTRF),urine microalbumin (UMA),urinary immunoglobulin IgG (UIgG),urine beta2-microglobulin (Uβ2-MG),Uα1-MG,urinary N-acetyl-beta-D-glucosaminidase (UNAG),plasma cystatin C (CysC) and Scr were detected by scattering turbidimetry,immune turbidimetry and fully automatic biochemical analysis pre-surgery 24 h and post-surgery 72 h.Contrast-induced acute kidney injury (CI-AKI) was defined as laboratory increase of Scr value≥44.2 μmol/L or ≥25％ from baseline measurement at 48 hours after surgery.The relationship in ioversol dosage and various factors was assessed by Single and multiple factors binary logistic regression analysis.Results According to the criterion that Scr increase value were ≥44.2 μmol/L,of 158 cases,3 cases occurred CI-AKI,the AKI incidence was 1.90％.Based on the criterion that Scr increase value was ≥25％,33 cases occurred CI-AKI,the incidence was 20.89％.The concentration of U-κ,UTRF,Uα1-MG,UNAG and plasma CysC were significantly different in high dose group compared to low ioversol dose group (P ＜ 0.05),while the other biomarkers had no significant difference (P ＞ 0.05).Conclusions The contrast media-ioversol could lead to CI-AKI;when the dosage of ioversol was more than 200 ml one-time,the concentration of U-κ,UTRF,Uα1-MG,UNAG and plasma CysC increased significantly.U-κ,UTRF,Uα1-MG,UNAG and plasma CysC could predict the early renal injury in patients who undergoing CVI.The rise of U-κ,UTRF,Uα1-MG,UNAG and plasma CysC are related to the dosage of ioversol.Furthermore,possibility of kidney injury is significantly high when ioversol dosage is more than 200 ml one-time.