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 construct an evaluation index system of primary practice bases for standardized training of assistant general practitioners in Beijing.Methods:From September to December 2023, the evaluation index system was initially developed using the Context, Input, Process, and Product Evaluation Model (CIPP model), then 20 experts engaged in clinical teaching management and general practice standardized training base evaluation were invited for three rounds of Delphi expert consultation, and the weights of the indicators at all levels were determined using the Analytic Hierarchy Process.Results:The response rates of the three rounds of expert consultation were 100.0%, 95.0%, and 95.0% respectively. The degree of authority of the expert consultations was greater than 0.70. The index importance and feasibility coordination coefficients of three rounds of expert consultation were 0.123 and 0.100 ( P<0.001), 0.064 and 0.068 ( P>0.05), and 0.149 and 0.221 ( P<0.001), respectively, indicting the convergence of the experts′ opinions. The evaluation indexes of the primary practice bases for standardized training of assistant general practitioners in Beijing were finally constructed, including five first-level indexes, including qualification and conditions, teaching organization and system, teaching staff, operation management and quality control; 11 second-level indexes, including basic conditions, department setup, teaching facilities, teaching organization and teaching system; and 25 third-level indexes, including qualification, scale, essential departments, conditional departments and basic teaching facilities. The weights of the five first-level indexes were qualification and conditions (0.333), teaching organization and system (0.167), teaching staff (0.167), operation management (0.167), quality control (0.167). Among the 11 second-level indexes, the top five weights were basic conditions (0.163), department setup (0.104), teaching organization (0.084), teacher requirements (0.084), and basic practice rotation management (0.084). Among the 25 third-level indexes, the top five weights were qualification (0.083), quality control process (0.082), scale (0.081), essential departments (0.053), and conditional departments (0.051). Conclusion:The evaluation index system of primary practice bases for standardized training of assistant general practitioners in Beijing has been constructed in this study based on Delphi expert consultations.

With the population ageing, the number of elderly with noncommunicable diseases and functional disabilities is increasing. The assistive technology can improve the ability of older adults for daily living activities and reduce dependence on caregivers to facilitate home-based care for the elderly. However, the provision of assistive technology service in China is still in the early stage and lacks implementable model. This article introduces international experiences on the delivery of assistive technology service, and discusses the status quo and problems of assistive technology service in China, to provide insights for promoting assistive technology service in the primary health care.

OBJECTIVE: To clarify the preparation methods of four rat models of liver ischemia/reperfusion injury (IRI) and to determine a liver IRI animal model that is consistent with clinical conditions, has stable pathological and physiological injury, and is easy to operate. METHODS: A total of 160 male Sprague-Dawley (SD) rats were randomly divided into four groups using an interval grouping method: 70% IRI (group A), 100% IRI (group B), 70% IRI with 30% hepatectomy (group C), and 100% IRI with 30% hepatectomy (group D), with 40 rats in each group. Each model was further divided into sham operation group (S group) and ischemia groups of 30, 60, and 90 minutes, with 10 rats in each group. After surgery, the survival status and awakening time of the rats were observed, and the liver lobectomy weight, bleeding volume, and hemostasis time of groups C and D were recorded. Blood samples were collected by cardiac puncture after 6 hours of reperfusion for determination the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), blood urea nitrogen (BUN), serum creatinine (SCr), and γ-glutamyl transpeptidase (γ-GT) in the serum to assess liver and kidney function. Hematoxylin-eosin (HE) staining and immunohistochemical staining of macrophages were performed to analyze the liver tissue structure damage from a pathological perspective. RESULTS: Rats in group A exhibited earlier awakening and acceptable mental status, while rats in the other groups showed delayed awakening and poor mental status. The hemostasis time in group D was approximately 1 second longer than that in group C. The mortality of rats subjected to 60 minutes of 70% hepatic ischemia was 0. Compared to the sham operation group, rats in each experimental group showed significant increases in serum levels of AST, ALT, ALP, BUN, SCr, and γ-GT, indicating impaired liver and kidney function in the rat models of liver IRI. In groups A, B, and C, the 90-minute ischemia subgroup exhibited more pronounced elevation in AST, ALT, ALP, BUN, SCr, and γ-GT levels compared to the 30-minute ischemia subgroup [AST (U/L): group A, 834.94±56.73 vs. 258.74±18.33; group B, 547.63±217.40 vs. 277.67±57.92; group C, 930.38±75.48 vs. 640.51±194.20; ALT (U/L): group A, 346.78±25.47 vs. 156.58±13.25; group B, 408.40±138.25 vs. 196.80±58.60; group C, 596.41±193.32 vs. 173.76±72.43; ALP (U/L): group A, 431.21±34.30 vs. 315.95±15.64; group B, 525.88±62.13 vs. 215.63±17.31; group C, 487.53±112.37 vs. 272.46±92.33; BUN (U/L): group A, 18.35±5.63 vs. 14.32±2.30; group B, 30.21±4.55 vs. 17.41±8.14; group C, 20.50±3.64 vs. 15.93±3.22; SCr (U/L): group A, 27.47±8.91 vs. 22.37±5.66; group B, 43.60±15.57 vs. 36.80±7.95; group C, 63.81±20.24 vs. 42.47±7.03; γ-GT (U/L): group A, 15.64±3.57 vs. 6.82±1.48; group B, 9.28±1.91 vs. 5.62±1.21; group C, 10.98±3.18 vs. 5.67±1.10; all P < 0.05]. The 100% IRI 90-minute group and 100% IRI 90-minute group with 30% hepatectomy exhibited more pronounced increases in the above-mentioned indicators compared to the corresponding 70% IRI control group, indicating increased liver and kidney damage in rats subjected to combined blood flow occlusion and hepatectomy. HE staining showed clear liver tissue structure with intact and orderly arranged cells in the sham operation group, while the experimental groups exhibited cell structure damage, including cell rupture or collapse, cell swelling, nuclear pyknosis, deep cytoplasm staining, cell shedding, and necrosis. The interstitium showed infiltration of inflammatory cells. Immunohistochemical staining revealed a higher number of macrophages in the experimental groups compared to the sham operation group. CONCLUSIONS Four models of liver IRI in rat were successfully established. As the duration and severity of hepatic ischemia increased, liver cell ischemia worsened, leading to increased hepatocellular necrosis and exhibiting characteristic features of liver IRI. These models can effectively simulate liver IRI following liver trauma, with the group subjected to 100% ischemia and 30% hepatectomy showing the most severe liver injury. The designed models are reasonable, easy to perform, and exhibit good reproducibility. They can be used for investigating the mechanisms, therapeutic efficacy, and diagnostic methods related to clinical liver IRI.
Rats ; Male ; Animals ; Reproducibility of Results ; Rats, Sprague-Dawley ; Liver ; Reperfusion Injury/drug therapy* ; Ischemia ; Disease Models, Animal ; Necrosis

Objective:To explore the effect of patient engagement on medication safety for patients with chronic disease through a systematic review.Methods:Relevant randomized controlled trials of patient engagement on medication safety were searched from PubMed, EMBASE, The Cochrane Library, CNKI, Wanfang and VIP database. The literature was screened according to inclusion and exclusion criteria, and data extraction and literature quality evaluation was conducted on the selected literature.Results:A total of 15 eligible studies was included. Most of the studies occurred in outpatient and home settings, and the subjects were patients with chronic diseases. Patient engagement strategies can be divided into three categories: (1) patient engagement in medical decision-making; (2) patient engagement in medication adjustment; (3) patient engagement in medication management. The outcomes of medication safety included medication adherence, medication knowledge, medication beliefs, adverse events and medication errors. Intervention strategies for patients to actively engage in medication safety significantly improved patients′ medication knowledge and beliefs, but did not improve medication adherence of patients.Conclusion:Promoting patient active engagement is an effective intervention measure to improve patients ′ perception of medication safety. Due to the limited quality and quantity of the included studies, more high-quality studies are needed to verify the above conclusions.

Objective:To compare gene expression profiles in normal human cervical epithelial cells (HcerEpic) before and after Chlamydia trachomatis ( Ct) infection. Methods:HcerEpic cells that were pretreated with DEAE-D were infected with Ct serotype E standard strain and then cultured for 44 h. Uninfected HcerEpic cells were used as the control group. Total RNA was extracted from the cells in each group and reverse transcribed to construct a cDNA library. Differences in gene expression profiles between the two groups were analyzed by high-throughput sequencing and the representative genes were selected for verification by qPCR. Results:A total of 23 997 genes were detected, including 125 differentially expressed genes. Among the 125 genes, 119 were up-regulated and six were down-regulated. GO analysis showed that the differentially expressed genes were enriched in several biological processes including defense response to virus, typeⅠinterferon signaling pathway and cellular responses to typeⅠinterferons. KEGG enrichment analysis showed the differentially expressed genes were mainly enriched in the pathways related to virus infections, such as influenza A virus, herpes simplex virus, EB virus and HPV, and NOD-like receptor pathway.Conclusions:There were significant differences in transcriptome profiles of HcerEpic cells before and after Ct infection. The differentially expressed genes were mainly enriched in the interferon pathway, which was closely related to the antiviral processes in cells. qPCR verified the differentially expressed genes and the genes closely related to the interferon pathway, such as ISG15, IFIT2, OASL and UBE2L6.

Cornelia de Lange syndrome (CdLS) is a genetic syndrome with severe neurodevelopmental disorders as the main manifestation. Its clinical manifestations included mental retardation, typical facial features, intrauterine and postnatal developmental delay, and deformity in multiple organs and systems, with an incidence rate of about 1/10000 to 1/30000. International CdLS Consensus Group was established in 2017 and issued the first international consensus on CdLS, i.e., "Diagnosis and management of Cornelia de Lange syndrome: first international consensus statement", in July 2018. Being developed through a modified Delphi consensus process, this consensus provides guidance on the diagnosis and management of children with CdLS. This article gives an interpretation of this consensus, aiming to help clinicians with early identification, diagnosis, standard follow-up, and management of this disease.
Consensus ; De Lange Syndrome ; Humans

Objective:To observe the clinical effect of prolonged photodynamic therapy (PDT) irradiation time combined with intravitreal injection of ranibizumab in the treatment of circumscribed choroidal hemangioma (CCH).Methods:A retrospective clinical study. From March 2012 to March 2018, 51 eyes of 51 patients diagnosed in Shenzhen Eye Hospital were included in the study. Among the patients, the tumor of 36 eyes were located in macular area, of 15 eyes were located outside macular area (near center or around optic disc). All patients underwent BCVA, color fundus photography, FFA, ocular B-scan ultrasonography and OCT examinations. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. OCT showed 48 eyes with macular serous retinal detachment. of 36 eyes with tumor located in macular area, the logMAR BCVA was 0.05±0.05, the tumor thickness was 4.5± 2.2 mm, the diameter of tumor was 9.7±3.6 mm. Of 15 eyes with tumor located outside macular area, the logMAR BCVA was 0.32±0.15, the tumor thickness was 3.8±1.4 mm, the diameter of tumor was 7.7± 1.9 mm. PDT was performed for all eyes with the irradiation time of 123 s. After 48 h, all patients received intravitreal injections of 0.5 mg ranibizumab (0.05 ml). At 1, 3 and 6 months after treatment, the same equipment and methods before treatment were used for related examination. BCVA, subretinal effusion (SRF), tumor leakage and size changes were observed. BCVA, tumor thickness and diameter before and after treatment were compared by t test. Results:At 6 months after treatment, the tumor was becoming smaller without scar formation. FFA showed that the blood vessels in the tumor were sparse compared with those before treatment, and the fluorescence leakage domain was reduced. OCT showed 43 eyes of macular serous detachment were treated after the combined treatment. The logMAR BCVA were 0.16±0.15 and 0.55±0.21 of the eyes with tumor located in or outside macular area, respectively. The difference of logMAR BCVA between before and after treatment was significant ( t=-2.511, -2.676; P=0.036, 0.040). Both the tumor thickness ( t=3.416, 3.055; P=0.011, 0.028) and diameter ( t=4.385, 4.171; P=0.002, 0.009) of CCH patients were significantly reduced compared with that before treatment. Conclusion:The tumor of CCH can be reduced by prolonged PDT irradiation time combined with intravitreal injection of ranibizumab.

OBJECTIVETo investigate the effects of siRNAs targeting CD97 immune epitopes on proliferation, infiltration, apoptosis and cell cycle of breast cancer cells.

METHODSsiRNA sequences targeting CD97and CD97immune epitopes were designed according to Gene Bank NM_001025160.2 with smart siCatchsiRNA design software. CD97siRNAs were transfected into MDA-MB231 cells in which CD97 was highly expressed. Highest sensitive CD97and CD97siRNA were screened by Western blotting. Inverted microscope was used to observe the growth of CD97siRNAs-transfected MDA-MB231 cells; the proliferation activity of MDA-MB231 cells was detected by MTT method; the wound healing assay and Transwell migration test were performed to examine the migration and infiltration ability of CD97and CD97siRNA-transfected MDA-MB231 cells; the effects of CD97siRNA and CD97siRNA on cell apoptosis and cell cycle of MDA-MB231 cells were detected by TUNEL and flow cytometry.

RESULTSThe growth and proliferation activity of CD97siRNAs-transfected MDA-MB231 cells were significantly lower than those in the control groups, and such differences were more significant in CD97siRNA-transfected group (all<0.05); scratch test showed that the wound healing rate was lower in CD97siRNAs-transfected groups, especially in CD97siRNA-transfected group (all<0.05); Transwell migration showed that the number of MDA-MB231 cells crossing through chambers were less in CD97siRNAs-transfected groups, especially in CD97siRNA-transfected group (all<0.05); no significant difference in cell apoptosis was observed between CD97siRNAs-transfected groups and control groups; cell cycle detection showed that CD97siRNAs-transfected groups had less cells in G/Gphase and more cells in S phase compared with the control groups, and such effect on cell cycle was more marked in CD97siRNA-transfected group (all<0.05).

CONCLUSIONSCD97 plays an important role in the cell growth, proliferation, migration and invasion of breast cancer MDA-MB231 cells, and compared with CD97, CD97may have more effective inhibitory effects on cellular malignant behaviors.

Objective To explore the clinical features of systemic lupus erythematosus (SLE) combined with antiphospholipid syndrome (APS) and cerebral thrombosis in a child. Method The clinical data of SLE combined with APS and cerebral thrombosis in a child was retrospectively analyzed, and the related literature was reviewed. Results This was a 12-year-old boy. The disease onset with recurrent fever, confusion and rash in cheek. He had anemia and thrombocytopenia, and positive antinuclear antibody (ANA) and anticardiolipin antibody (aCL). Magnetic resonance imaging showed multiple cerebral infarction. The diagnosis of SLE combined with APS and cerebral thrombosis was clearly made. Methylprednisolone, cyclophosphamide, warfarin, meropenem and acyclovir were used for the treatment. At the same time, the patient also received intravenous immunoglobulin. Conclusion SLE combined with APS and cerebral thrombosis in children was usually in a severe condition, the prognosis of which can be effectively improve by early diagnosis and reasonable treatment.