Objective To investigate the basic information of human immunodeficiency virus/acquired immune deficiency syndrome(HIV/AIDS)patients who infected with Epstein-Barr virus(EBV)or human Cytomegalovirus(HCMV),collect the relevant clinical immunological data and analyze the influencing factors.Method A total of 1 093 HIV/AIDS patients treated in the First Hospital of Changsha from January to December 2022 and underwent EBV and HCMV screening were collected.Flow cytometry was used to detect the CD4+T lymphocytes.Fluorescence quantitative PCR was applied for HIV-RNA,EBV-DNA,and HCMV-DNA testing.Statistical analysis was carried out by using SPSS 27.0,and logistic regression was used to analyze the risk factors of HIV/AIDS patients complicated with viral infection.Results Among 1 093 HIV/AIDS patients,the positive rates of EBV-DNA and HCMV-DNA were 48.22%(527/1 093)and 19.03%(208/1 093),respectively.As the number of CD4+T lymphocytes increased,the positive rates of EBV-DNA and HCMV-DNA decreased,and the differences was statistically significant(χ2=39.50,143.0,all P<0.001).As the level of HIV-RNA increased,the positive rates of EBV-DNA and HCMV-DNA increased,and the differences were statistically significant(χ2=46.18,124.3,all P<0.001).The patients receiving antiretroviral therapy(ART)significantly decreased the positive rates of EBV-DNA and HCMV-DNA(χ2=30.60,96.59,all P<0.001).There was a significant negative correlation between the number of CD4+T lymphocytes and the level of HIV-RNA(r=-0.49,P<0.001).Logistic regression analysis showed that the CD4+T lymphocyte count<200/μl(OR=1.46,95%CI:1.02～2.08,P=0.037),HIV-RNA load>200 copies/ml(OR=1.70,95%CI:1.18～2.44,P=0.004)and the age>30 years old(OR=2.15,95%CI:1.44～3.19,P<0.001)were risk factors for HIV/AIDS patients infected with EBV.Without regularly receiving ART(OR=1.83,95%CI:1.10～3.02,P=0.019),HIV-RNA load>200 copies/ml(OR=2.56,95%CI:1.50～4.35,P<0.001)and the CD4+T lymphocyte count<200/μl(OR=4.61,95%CI:2.57～8.28,P<0.001)were risk factors for HCMV infection in HIV/AIDS patients.Conclusion To reduce the possibility of opportunistic infection in HIV/AIDS patients,the surveillance of EBV and HCMV and regular ART should be strengthened,especially when the number of CD4+T lymphocytes decreases(<200/μl),the level of HIV RNA increases(>200 copies/ml)or the age>30 years old.

Clinical trials are an important method for evaluating the safety and efficacy of in vitro diagnostic reagents, and are a key basis for product registration review and approval. In order to strengthen the management of clinical trials of in vitro diagnostic reagents, the National Medical Products Administration and relevant departments have formulated a series of regulations at the regulatory level, and require applicants and clinical trial institutions to establish a quality management system for clinical trials of in vitro diagnostic reagents. Medical laboratory is the main department and implementer of in vitro diagnostic reagent clinical trials in medical institutions. In recent years, with the rapid development of the in vitro diagnostic industry, the clinical trial projects of in vitro diagnostic reagents conducted by medical laboratory have been increasing day by day. However, there are currently few discussions on the clinical trial of in vitro diagnostic reagents from the perspective of researchers. Therefore, this article summarizes the characteristics of clinical trials of in vitro diagnostic reagents, analyzes the problems and difficulties in conducting clinical trials of in vitro diagnostic reagents in current medical laboratories, and introduces the laboratory′s experience in management; to provide reference for medical testing laboratories that have not yet conducted or have already conducted clinical trials of in vitro diagnostic reagents, in order to improve the quality and efficiency of clinical trials.

Clinical trials are an important method for evaluating the safety and efficacy of in vitro diagnostic reagents, and are a key basis for product registration review and approval. In order to strengthen the management of clinical trials of in vitro diagnostic reagents, the National Medical Products Administration and relevant departments have formulated a series of regulations at the regulatory level, and require applicants and clinical trial institutions to establish a quality management system for clinical trials of in vitro diagnostic reagents. Medical laboratory is the main department and implementer of in vitro diagnostic reagent clinical trials in medical institutions. In recent years, with the rapid development of the in vitro diagnostic industry, the clinical trial projects of in vitro diagnostic reagents conducted by medical laboratory have been increasing day by day. However, there are currently few discussions on the clinical trial of in vitro diagnostic reagents from the perspective of researchers. Therefore, this article summarizes the characteristics of clinical trials of in vitro diagnostic reagents, analyzes the problems and difficulties in conducting clinical trials of in vitro diagnostic reagents in current medical laboratories, and introduces the laboratory′s experience in management; to provide reference for medical testing laboratories that have not yet conducted or have already conducted clinical trials of in vitro diagnostic reagents, in order to improve the quality and efficiency of clinical trials.

Objective:High-throughput screening to obtain small molecular compounds against Gram-negative bacilli by targeting BamA outer membrane protein.Methods:The sybyl-X2.1 software was used to perform high-throughput virtual screening of small molecular compounds in Chemdiv compound library based on the molecular docking. The top 150 hits by high-throughput screening were re-screened through in vitro biological experiments. The top 4 small molecules with obvious antibacterial activity were selected for in-depth molecular docking analysis, and the small molecule 8308-0401 with the highest docking score was selected for further experiments. The antibacterial effect of 8308-0401 combined with rifampicin was tested by checkerboard assay. Finally, the affinity between 8308-0401 and BamA was tested by plasma surface resonance assay. Results:The docking score of the top 150 hits calculated by high-throughput virtual screening had a mean value of 5.63. In vitro biological experiments showed that small molecules 8308-0401, 8365-1335, C066-2507 and L582-0346 exhibited strong antibacterial activity. Among those molecules, 8308-0401 showed the highest molecular docking score, and synergistic antibacterial activity against both types of strains and clinical isolates when combined with rifampicin. 8308-0401 has a strong affinity to BamA with binding a constant of 182 μmol/L. Conclusion:The small molecule 8308-0401 exerts antibacterial activity against Gram negative bacilli by targeting the outer membrane protein BamA.

Objective:To investigate the clinical characteristics and possible causes of transient spontaneous remission of childhood acute leukemia.Methods:The data of 3 children with acute leukemia who had transient spontaneous remission before standardized chemotherapy in Sun Yat-sen Memorial Hospital of Sun Yat-sen University in July 2018, May 2019 and October 2020 were collected. Moreover, the related influencing factors of spontaneous remission in leukemia were discussed by review of the literature.Results:All 3 children had fever at the onset of the disease, and they achieved transient spontaneous remission after anti-infection therapy. Case 1 obtained partial remission after the initial diagnosis of acute B lymphocytic leukemia (B-ALL), leukemia gene test showed E2A-PBX1 fusion, and relapsed after 12 days. Case 2 obtained spontaneous remission after the initial diagnosis of B-ALL, leukemia gene test showed p16 gene deletion and NRAS and EP300 genes mutation, and relapsed after 20 days. Case 3 obtained spontaneous remission after the initial diagnosis of acute monocytic leukemia, leukemia gene test showed MLL-ENL fusion and NRAS gene mutation, and relapsed after 30 days. A review of the literature showed that the main influencing factors of spontaneous remission in leukemia were Down syndrome, infection and blood transfusion. Other influencing factors included leukemia-related genes, termination of pregnancy and application of drugs.Conclusions:Transient spontaneous remission of childhood acute leukemia is rare in clinical practice, and the possible mechanism is related to infection-induced immune abnormalities. It is recommended that leukemia patients with spontaneous remission should be closely monitored for minimal residual disease.

Objective:To analyze the clinical effect of hyaluronidase injection through the facial artery in the treatment of vascular embolism such as skin ulceration and necrosis after cosmetic injection.Methods:Hyaluronidase was injected through facial artery in 13 patients who were diagnosed with vascular embolism after facial injection from January 2019 to May 2020. The facial artery was punctured with 22-gauge arterial blood collection needle or 19/23-gauge disposable venous infusion needle. The angle between the needle body and the skin varies depending on the patients’ weight, ranged 30°-45°. The needle was advanced slowly and pushed forward by 2-3 mm when blood backflow appeared in the needle core. After confirming the successful puncture of the facial artery, 0.5-1.5 ml hyaluronidase was slowly injected into the facial artery. The time of skin relaxation, tenderness relief, ulcer healing and wound recovery were observed. The pigmentation was observed and the Vancouver Scar Scale (VSS) was used to score the scars after 3-12 months.Results:A toal of 13 patients with vascular complications of hyaluronidase filler were retrospectively reviewed. The patients were 18-45-year-old(mean age, 35 years) and received hyaluronidase filler at private clinics. There were 12 women and 1 man. The time from onset to visit was 14 h to 4 d, with an average time of 2.5 d. Hyaluronidase was most commonly injected into the nasolabial folds (54%, 7 of 13). The second-ranked area is the nasalroot (23%, 3 of 13). These patients had skin swelling, necrosis, ecchymosis or black scabs during or after hyaluronidase injection. Some patients showed skin lesions combined with oral ulcer. After percutaneous facial arterial hyaluronidase injection, the local skin tissue injuries of the 13 patients were improved in time. The time of skin relaxation was (0.77±0.25) d, the time of tenderness relief was (1.23±0.64) d, the time of ulcer healing was (3.14±0.64) d and the time of wound recovery was (5.85±0.86) d. Patients were followed up for 3-12 months, with an average of 7 months. One patient had slight scar (VSS score of 1), two patients had only mild pigmentation (VSS score of 0), and the other ten patients had no scar and pigmentation (VSS score of 0).Conclusions:It is effective to improve local microcirculation and reduce skin tissue injury after percutaneous facial artery hyaluronidase injection in the treatment of skin injury caused by facial filler injection.

Objective:To analyze the clinical effect of hyaluronidase injection through the facial artery in the treatment of vascular embolism such as skin ulceration and necrosis after cosmetic injection.Methods:Hyaluronidase was injected through facial artery in 13 patients who were diagnosed with vascular embolism after facial injection from January 2019 to May 2020. The facial artery was punctured with 22-gauge arterial blood collection needle or 19/23-gauge disposable venous infusion needle. The angle between the needle body and the skin varies depending on the patients’ weight, ranged 30°-45°. The needle was advanced slowly and pushed forward by 2-3 mm when blood backflow appeared in the needle core. After confirming the successful puncture of the facial artery, 0.5-1.5 ml hyaluronidase was slowly injected into the facial artery. The time of skin relaxation, tenderness relief, ulcer healing and wound recovery were observed. The pigmentation was observed and the Vancouver Scar Scale (VSS) was used to score the scars after 3-12 months.Results:A toal of 13 patients with vascular complications of hyaluronidase filler were retrospectively reviewed. The patients were 18-45-year-old(mean age, 35 years) and received hyaluronidase filler at private clinics. There were 12 women and 1 man. The time from onset to visit was 14 h to 4 d, with an average time of 2.5 d. Hyaluronidase was most commonly injected into the nasolabial folds (54%, 7 of 13). The second-ranked area is the nasalroot (23%, 3 of 13). These patients had skin swelling, necrosis, ecchymosis or black scabs during or after hyaluronidase injection. Some patients showed skin lesions combined with oral ulcer. After percutaneous facial arterial hyaluronidase injection, the local skin tissue injuries of the 13 patients were improved in time. The time of skin relaxation was (0.77±0.25) d, the time of tenderness relief was (1.23±0.64) d, the time of ulcer healing was (3.14±0.64) d and the time of wound recovery was (5.85±0.86) d. Patients were followed up for 3-12 months, with an average of 7 months. One patient had slight scar (VSS score of 1), two patients had only mild pigmentation (VSS score of 0), and the other ten patients had no scar and pigmentation (VSS score of 0).Conclusions:It is effective to improve local microcirculation and reduce skin tissue injury after percutaneous facial artery hyaluronidase injection in the treatment of skin injury caused by facial filler injection.

Objective:To investigate the expression of oxidative stress and related pathway factors in fibroblasts from human hypertrophic scar and normal skin tissue.Methods:Select and collect the scar tissue and normal full-thickness skin tissue of the donor area from 8 hypertrophic scar patients who were treated with surgical resection at the Department of Burns and Plastic Surgery, the Fourth Medical Center of Chinese PLA General Hospital from June 2019 to July 2020, extract and culture primary fibroblasts by weaving method, subculture them, and perform the following experiments when the cells were subcultured to the third generation: (1)Detect the proliferation ability of hypertrophic scar fibroblasts(HSF)and normal skin fibroblasts(NSF)with CCK-8 method.(2)Detect the number of reactive oxygen species in HSF and NSF by flow cytometry.(3)Colorimetric method for detecting relative activity of superoxide dismutase(SOD)in two kinds of cells.(4)Detect the NQO1 gene expression in two kinds of cells with RT-PCR.(5)Detect the Nrf2 gene expression in two kinds of cells with RT-PCR.(6)Detects Nrf2 and Bcl2 protein content in two kinds of cells with Western blotting.(7)Detect the expression and distribution of Nrf2 in cells with cellular immunohistochemical staining. The results were analyzed by SPSS 25.0 statistical software, independent sample t test was performed, and P<0.05 was considered as the difference was statistically significant. Results:(1) Compared with NSF, the number of the two kinds of cells is statistically different from the third day (The statistical results from the third day to the seventh day were as follows: t=2.631, P=0.039; t=7.025, P<0.001; t=5.031, P<0.001; t=4.241, P=0.002; t=6.525, P=0.003). (2) The reactive oxygen species content in HSF was 75.39%±3.06%, which was significantly higher than 45.36%±3.66% in the NSF group ( t=-17.804, P<0.001). (3) The relative activity of SOD of the HSF group was (50.76±0.52) U/ml, which is higher than that of the NSF group (42.76±1.35) U/ml ( t=15.674, P<0.001). (4) The relative expression of NQO1 mRNA in HSF was 0.859±0.076, which was lower than that in the NSF group at 1.595±0.181 ( t=3.763, P=0.020). (5) The relative expression of Nrf2 mRNA in HSF was 0.590±0.055, which was lower than that in the NSF group at 1.595±0.146 ( t=6.445, P=0.003). (6) The relative expression of Nrf2 protein of HSF was 0.314±0.035, which was significantly lower than 0.912±0.039 in the NSF group ( t=22.554, P<0.001). The relative expression of Bcl2 protein of HSF was 0.466±0.020, which was significantly lower than 0.734±0.066 in the NSF group ( t=7.780, P<0.001). (7) Nrf2 protein express in NSF and HSF cells, and protein expression was found in both nucleus and cytoplasm. Conclusions:HSF has a high degree of oxidative stress, which may be due to some reasons that reduce the content of Nrf2, resultsing in a decrease in the expression of various antioxidant enzymes, and ultimately leading to the accumulation of reactive oxygen species. Compared with NSF, HSF has stronger proliferation and apoptosis abilities, and reactive oxygen species can cause cell apoptosis, indicating that oxidative stress may be one of the pathogenesis of hypertrophic scars.

Objective:To investigate the expression of oxidative stress and related pathway factors in fibroblasts from human hypertrophic scar and normal skin tissue.Methods:Select and collect the scar tissue and normal full-thickness skin tissue of the donor area from 8 hypertrophic scar patients who were treated with surgical resection at the Department of Burns and Plastic Surgery, the Fourth Medical Center of Chinese PLA General Hospital from June 2019 to July 2020, extract and culture primary fibroblasts by weaving method, subculture them, and perform the following experiments when the cells were subcultured to the third generation: (1)Detect the proliferation ability of hypertrophic scar fibroblasts(HSF)and normal skin fibroblasts(NSF)with CCK-8 method.(2)Detect the number of reactive oxygen species in HSF and NSF by flow cytometry.(3)Colorimetric method for detecting relative activity of superoxide dismutase(SOD)in two kinds of cells.(4)Detect the NQO1 gene expression in two kinds of cells with RT-PCR.(5)Detect the Nrf2 gene expression in two kinds of cells with RT-PCR.(6)Detects Nrf2 and Bcl2 protein content in two kinds of cells with Western blotting.(7)Detect the expression and distribution of Nrf2 in cells with cellular immunohistochemical staining. The results were analyzed by SPSS 25.0 statistical software, independent sample t test was performed, and P<0.05 was considered as the difference was statistically significant. Results:(1) Compared with NSF, the number of the two kinds of cells is statistically different from the third day (The statistical results from the third day to the seventh day were as follows: t=2.631, P=0.039; t=7.025, P<0.001; t=5.031, P<0.001; t=4.241, P=0.002; t=6.525, P=0.003). (2) The reactive oxygen species content in HSF was 75.39%±3.06%, which was significantly higher than 45.36%±3.66% in the NSF group ( t=-17.804, P<0.001). (3) The relative activity of SOD of the HSF group was (50.76±0.52) U/ml, which is higher than that of the NSF group (42.76±1.35) U/ml ( t=15.674, P<0.001). (4) The relative expression of NQO1 mRNA in HSF was 0.859±0.076, which was lower than that in the NSF group at 1.595±0.181 ( t=3.763, P=0.020). (5) The relative expression of Nrf2 mRNA in HSF was 0.590±0.055, which was lower than that in the NSF group at 1.595±0.146 ( t=6.445, P=0.003). (6) The relative expression of Nrf2 protein of HSF was 0.314±0.035, which was significantly lower than 0.912±0.039 in the NSF group ( t=22.554, P<0.001). The relative expression of Bcl2 protein of HSF was 0.466±0.020, which was significantly lower than 0.734±0.066 in the NSF group ( t=7.780, P<0.001). (7) Nrf2 protein express in NSF and HSF cells, and protein expression was found in both nucleus and cytoplasm. Conclusions:HSF has a high degree of oxidative stress, which may be due to some reasons that reduce the content of Nrf2, resultsing in a decrease in the expression of various antioxidant enzymes, and ultimately leading to the accumulation of reactive oxygen species. Compared with NSF, HSF has stronger proliferation and apoptosis abilities, and reactive oxygen species can cause cell apoptosis, indicating that oxidative stress may be one of the pathogenesis of hypertrophic scars.

Objective To explore the diagnosis and treatment of non-tuberculosis mycobacteria (NTM) infection after cosmetic injection via scientific debridement surgery combined with regular application of anti-NTM drugs.Methods 14 patients who were infected with NTM after cosmetic injection and were not cured over a long period of time in other hospitals from 2012 to 2016.The patients were treated with VSD thorough surgical debridement,the bacterial type of NTM was identified by bacterial culture and PCR identification and anti-NTM drugs were systematically used according to the results of drug sensitivity.Results Fourteen patients who were treated with scientific debridement surgeries combined with regular anti-NTM drug treatment in our hospital for 2-4 months were discharged after their skin lesions were cleared and healed and they were continually treated with antiNTM drugs for 12 months.Fourteen patients were completely cured by using the above treatments without severe side effects,such as liver and kidney dysfunction,nervous system disorders and so on.Only colpitis mycotica occurred in 3 patients.In addition,one patient presented the decrease of leukopenia after using anti-NTM drugs for 2 months and continued to complete the treatment after we adjusted the treatment plan to returning the level of leukopenia to the normal.These 14 patients were followed up for 1-5 years with no recurrence of the lesion.The facial appearance of 12 patients were almost normal with slight scars.The facial surgery area of 2 patients were uneven and nearly recovered to normal facial appearance by tissue transplantation and photoelectric therapy.Conclusions For the NTM patients caused by invasive procedures such as injection,the comprehensive treatment program,which combined scientific debridement surgery and systematically targeted drug treatment,not only can effectively cure NTM infection,but also minimize secondary injury and restore the patients' appearance,which is worthy of clinical application.