[Objective] To extract hemoglobin (Hb) from red blood cells using osmotic pressure swelling method, expected to achieve a hemoglobin dissolution rate of ≥80% and a cell membrane integrity rate of ≥70%. [Methods] Human umbilical cord blood red blood cells were used as raw materials and phosphate buffer solution was used as the swelling solution for red blood cells. A three factor three-level orthogonal experiment (n=3) was conducted to determine the optimal matching conditions for selecting the osmolality molar concentration of phosphate buffer solution, pH value of hypotonic phosphate buffer solution and volume ratio of hypotonic phosphate buffer solution to washed red blood cells. Red blood cell swelling solution samples (n=6) were prepared by the optimal matching conditions and the original process conditions. The hemoglobin dissolution rate and cell membrane integrity rate were checked. In the expanded comparative experiment, red blood cell swelling solution samples (n=6) were prepared by the optimal matching conditions and the original process conditions, which was filtered by ultrafiltration membranes. The filtration time and hemoglobin yield were checked. [Results] The optimal matching conditions for preparing red blood cell swelling solution were obtained through orthogonal experiment as follows: osmotic pressure molar concentration was 30 mOsmol/Kg, pH was 7.8, and phosphate buffer to red blood cell volume ratio was 6∶1. On the basis of the above conditions, the red blood cell swelling solution sample was compared with the original process sample: the hemoglobin dissolution rate was (82.4±1.8)% vs (78.6±3.0)% (P<0.05), and the cell membrane integrity rate was (65.8±4.0)% vs (28.7±2.3)% (P<0.05). In the expanded comparative experiment, the optimal matching conditions were compared with the original process conditions: filtration time(s) (327±9) vs (434±13) (P<0.05), and hemoglobin yield was (72.3±1.2)% vs (66.0±1.4)% (P<0.05). [Conclusion] Compared with the original preparation process, the hemoglobin extraction process which optimized through orthogonal experiments greatly reduces the cell membrane fragmentation rate and minimizes the entry of cell membrane matrix into the target solution, ensuring a slightly higher hemoglobin dissolution rate, and reducing the preparation difficulty for the subsequent cell membrane separation and further purification.

Objective: To study the effect of nano hemoglobin-based oxygen carrier (nano-HBOC) on radiosensitivity of lung cancer H385 cells. Methods: Using 95% N and 5% CO , a lung cancer cell line was constructed in a hypoxic environment, and H385 cells were treated with different concentrations of nano-HBOC and irradiated (4Gy) by an irradiator, and the IC50 concentration was calculated. The cells were detected by flow cytometry (reactive oxygen species, ROS) ROS test. Using GEO database, KEGG pathway enrichment analysis was carried out to predict possible pathways. The levels of lipid peroxidation and Fe were observed by fluorescence microscope, and the proteins related to iron death pathway were detected by Western-blot. Results: Compared with the control cells, the activity and density of the cells were significantly decreased by nano-HBOC combined with radiotherapy, with a notable proportion of cells exhibiting deteriorated status. There is a positive correlation between ROS level and nano-HBOC concentration, especially after radiotherapy. Radiotherapy combined with nano-HBOC significantly increased the levels of lipid peroxidation and Fe in H385 cells, while decreasing the levels of iron death pathway proteins slc7a11 and GPX4, and increasing the level of ACSL4. Conclusion: Nano-HBOC enhances the radiosensitivity of lung cancer H385 cells.

Objective: To study the effect of nano hemoglobin-based oxygen carrier (nano-HBOC) on radiosensitivity of lung cancer H385 cells. Methods: Using 95% N and 5% CO , a lung cancer cell line was constructed in a hypoxic environment, and H385 cells were treated with different concentrations of nano-HBOC and irradiated (4Gy) by an irradiator, and the IC50 concentration was calculated. The cells were detected by flow cytometry (reactive oxygen species, ROS) ROS test. Using GEO database, KEGG pathway enrichment analysis was carried out to predict possible pathways. The levels of lipid peroxidation and Fe were observed by fluorescence microscope, and the proteins related to iron death pathway were detected by Western-blot. Results: Compared with the control cells, the activity and density of the cells were significantly decreased by nano-HBOC combined with radiotherapy, with a notable proportion of cells exhibiting deteriorated status. There is a positive correlation between ROS level and nano-HBOC concentration, especially after radiotherapy. Radiotherapy combined with nano-HBOC significantly increased the levels of lipid peroxidation and Fe in H385 cells, while decreasing the levels of iron death pathway proteins slc7a11 and GPX4, and increasing the level of ACSL4. Conclusion: Nano-HBOC enhances the radiosensitivity of lung cancer H385 cells.

By summarizing the biological formation and component function of Mycobacterium tuberculosis extracellular vesicles, this manuscript discusses the potential usage of Mycobacterium tuberculosis extracellular vesicles in diagnosis, prevention, and treatment of tuberculosis. It is concluded that both Mycobacterium tuberculosis membrane vesicles and exosomes secreted by the host contain the components of Mycobacterium tuberculosis to participate in immune regulation, and play different roles in the diagnosis, prevention, and treatment of tuberculosis. Mycobacterium tuberculosis extracellular vesicles contain abundant biomarkers and have natural immunogenicity, which are expected to become new targets for diagnosis, prevention, and treatment of tuberculosis.

Objective:To analyze the short-term clinical efficacy and safety of arsenic trioxide (ATO) combined with a modified N7 induction regimen in the treatment of children with high-risk neuroblastoma (NB).Methods:This study was a prospective, single-arm, multicenter phase Ⅱ clinical study. Sixty-seven high-risk NB children from eight units of Sun Yat-sen Memorial Hospital of Sun Yat-sen University, Wuhan Children′s Hospital of Tongji Medical College of Huazhong University of Science and Technology, First Affiliated Hospital of Guangxi Medical University, Hainan General Hospital, Affiliated Hospital of Guangdong Medical University, Kunming Children′s Hospital, Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology, and Guangdong Provincial Agricultural Reclamation Center Hospital were enrolled from January 2019 to August 2023 and were treated with ATO combined with a modified N7 induction regimen. The efficacy and adverse effects at the end of induction chemotherapy were assessed and analyzed, and the differences in the clinical characteristics were further compared between the treatment-responsive and treatment-unresponsive groups by using the Fisher′s exact test.Results:Among 67 high-risk NB children, there were 40 males (60%) and 27 females (40%), with the age of disease onset of 3.5 (2.6, 4.8) years. Primary NB sites were mostly in retroperitoneum (including adrenal gland) (56/67, 84%) and the common metastases sites at initial diagnosis were distant lymph node in 25 cases (37%),bone in 48 cases (72%),bone marrow in 56 cases (84%) and intracalvarium in 3 cases (4%). MYCN gene amplification were detected in 28 cases (42%). At the end of induction, 33 cases (49%) achieved complete remission, 29 cases (43%) achieved partial remission, 1 case (1%) with stable disease, and 4 cases (6%) were assessed as progressive disease (PD). The objective remission rate was 93% (62/67) and the disease control rate was 94% (63/67). The percentage of central system metastases at the initial diagnosis was higher in the treatment-unresponsive group than in the treatment-responsive group (2/5 vs. 2% (1/62), P=0.013), whereas the difference in MYCN gene amplification was not statistically significant between two groups (3/5 vs.40% (25/62), P=0.786). Grade Ⅲ or higher adverse reactions during the induction chemotherapy period were myelosuppression occurred in 60 cases (90%), gastrointestinal symptoms occurred in 33 cases (49%), infections occurred in 20 cases (30%), hepatotoxicity occurred in 4 cases (6%), and cardiovascular toxicity occurred in 1 case (2%). There were no chemotherapy-related deaths. Conclusion:ATO combined with N7-modified induction regimen had a superiority in efficacy and safety, which deserved further promotion in clinical practice.

Objective To explore the application value of different preprotein converting subtilisin/kexin type 9 (PCSK9) inhibitors in familial hypercholesterolemia(FH). Methods Patients with FH in our hospital were selected and divided into alirocumab group and evolocumab group according to the different PCSK9 inhibitors after excluding the confounding factors of baseline data such as gender and age by propensity score matching, 41 patients in each group were matched. Both groups were treated for 3 months. The therapeutic effect, and blood lipids levels[triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) index, apolipoprotein A1 (Apo A1), apolipoprotein B (Apo B)], coronary flow reserve[absolute coronary flow reserve (CFR), relative coronary flow reserve (rCFR), fractional flow reserve (FFR)], endothelial function indicators[serum nitric oxide (NO), endothelin-1 (ET-1), flow mediated dilatation (FMD)]and adverse reactions before and after treatment were compared between both groups. Results After three months of treatment, the overall standardized rate of LDL-C in the alirocumab group was 97.56 % (40/41), and 92.68 % in the evolocumab group, but there was no statistically significant difference between the two groups (P>0.05). After treatment, the levels of TG, TC, LDL-C, Apo B, and ET-1 in both groups decreased, while the levels of HDL-C, Apo A1, CFR, rCFR, FFR, NO, and FMD increased. Additionally, the levels of TG, TC, LDL-C, Apo B, and ET-1 in the alirocumab group were lower than those in the evolocumab group, while the levels of HDL-C, Apo A1, CFR, rCFR, FFR, NO, and FMD were higher, and the differences were statistically significant (P < 0.05). The incidence of adverse reactions was 12.20 % in the alirocumab group and 9.76 % in the evolocumab group, with no statistically significant difference (P>0.05). Conclusion Alirocumab or evolocumab combined with rosuvastatin in treating FH can effectively improve the lipid metabolism, coronary flow reserve function and vascular endothelial function in the treatment of patients with hypercholesterolemia, but alirocumab has a better effect.

Objective To explore the standardized management mode of the Ethics Committee for organ donation after citizen’s death in hospitals. Methods The situations of ethical review before and after the standardized adjustment of the Ethics Committee of human organ donation in the First Affiliated Hospital of Chongqing Medical University were retrospectively analyzed. Baseline data of donors before and after standardized adjustment of the Ethics Committee of human organ donation were compared. The influence of standardized adjustment of the Ethics Committee on the attendance rate of committee members and duration of ethical review were analyzed. Results No significant differences were observed in donors' ethical review data, such as gender, age and death determination, before and after standardized adjustment of Ethics Committee structure (all P>0.05). Significant difference was noted regarding the cause of death in ethical review (P<0.05). Univariate analysis showed that there were significant differences in the impact of Ethics Committee standardization adjustment and cause of death on the attendance rate of committee members (both P<0.05). Multivariate analysis revealed that gender, cause of death and standardized adjustment of the Ethics Committee were the influencing factors of the attendance rate of committee members, and the attendance rate of committee members after standardized adjustment was higher than that before adjustment (P<0.05). Univariate analysis showed that there were statistically significant differences in the effects of Ethics Committee standardized adjustment, attendance rate of committee members and cause of death on the duration of ethical review (all P<0.05). Multivariate analysis indicated that standardized adjustment of the ethics committee was the influencing factor of the duration of ethical review, and the duration of ethics review after standardized adjustment was shorter than that before adjustment (P<0.05). Conclusions Appropriate arrangement of the total number of ethics committee members and standardizing the review process may improve the efficiency of ethical review. Scientific evaluation mechanism for ethical committee members should be established by dynamically adjusting the ethical committee members, clarifying the responsibilities and tasks of members and secretaries, aiming to further improve standardized management level of ethical review for organ donation after citizen’s death.

Objective:To analyze the clinicopathological features in diabetic kidney disease (DKD) and non-diabetic kidney disease (NDKD) patients, and provide reference for patients who will receive renal biopsy with diabetes mellitus complicated with chronic kidney disease.Methods:The patients with type 2 diabetes mellitus complicated with chronic kidney disease who underwent renal biopsy were collected through the database at the Nanfang Hospital of Southern Medical University from February 2002 to June 2018. According to the results of renal biopsy, they were divided into DKD group and NDKD group (including DKD+NDKD). The clinical manifestations and pathological types were compared between the two groups.Results:A total of 507 patients were eventually included in the study. There were 114 cases (22.5%) with DKD and 393 cases (77.5%) with NDKD. Pathologically, the most common pathological types of NDKD were membranous nephropathy (30.0%) and IgA nephropathy (19.1%). Among NDKD patients, 5.6% patients had DKD combing with NDKD. In term of the clinical manifestations, DKD patients had a longer history of diabetes (>1 year, 76.3% vs 36.1%, P<0.001), higher quantity of urinary protein [3.69(1.70, 6.74) g/24 h vs 2.21(0.91, 4.97) g/24 h, P<0.001], higher serum creatinine [117.5(85.8, 194.5) μmol/L vs 89.0(68.0, 143.8) μmol/L, P<0.001] than NDKD patients. But the hemoglobin [(105.07±20.85) g/L vs (124.41±25.02) g/L, P=0.002] and cholesterol [(5.69±1.87) mmol/L vs (6.43±2.75) mmol/L, P=0.001] in DKD patients were lower than those in NDKD patients. Logistic regression analysis showed that diabetes mellitus history ( OR=4.162, 95% CI 1.717-10.098, P=0.002) , higer systolic pressure (every 1 mmHg, OR=1.028, 95% CI 1.011-1.045, P=0.001) , history of antihypertensive medication ( OR=3.141, 95% CI 1.496-6.591, P=0.002), diabetic retinopathy ( OR=5.561, 95% CI 2.361-13.100, P<0.001) and higher glycated hemoglobin level (every 1%, OR=1.680, 95% CI 1.333-2.118, P<0.001) were related factors of DKD, while hematuria ( OR=2.781, 95% CI 1.334-5.798, P=0.006) and higher hemoglobin level (every 1 g/L, OR=1.022, 95% CI 1.008-1.037, P=0.002) were related factors of NDKD. Conclusions:There are differences in clinical manifestations and pathological types between DKD and NDKD. The history of diabetes, antihypertensive medication, fundus examination, higher of proteinuria and glycosylated hemoglobin may predict DKD, while hematuria and higher level of hemoglobin may have certain guiding significance for the diagnosis of NDKD. The indication of renal biopsy in patients with diabetes mellitus complicated with chronic kidney disease should include comprehensive clinical manifestations.

Adult ; Child ; Female ; Humans ; Keratoderma, Palmoplantar ; genetics ; pathology ; Male ; Pedigree

Objective To investigate the distribution and drug resistance to pathogenic bacterial pathogen in children with community acquired pneumonia (CAP),so as to provide recommendations for clinical rational use of anti-biotics. Methods A retrospective analysis was made on the distribution and drug resistance to bacteria in CAP chil-dren admitted to Department of Respiration,Shanghai Children′s Medical Center from January 2014 to December 2015. Results There were 463 patients with positive sputum culture,and a total of 496 strains of pathogens were found. There were 273 Galanz negative bacteria,195 Galanz positive bacteria and 28 other rare bacteria,accounted for 55. 04％,39. 31％ and 5. 65％ of the total bacteria,respectively. The main pathogens were Streptococcus pneumoniae, Staphylococcus aureus,Haemophilus influenzae,Klebsiella pneumoniae and Escherichia coli. The highest detection rate of bacteria in 1-12 months children with CAP was Staphylococcus aureus,Klebsiella pneumoniae and Escherichia coli;in > 12 months children with CAP,the highest detection rate of bacteria was Streptococcus pneumoniae,Haemophilus influenzae and Staphylococcus aureus. Both of Streptococcus pneumoniae and Staphylococcus aureus had a high resis-tance to Erythromycin,Clindamycin and Oxacillin. There were 11. 00％ Streptococcus pneumoniae and 94. 74％ taphy-lococcus aureus resistant to Penicillin,while they were not resistant to Vancomycin. Escherichia coli and Klebsiella pneumoniae both showed a high resistance to ampicillin,the second and third generation cephalosporins. Haemophilus influenzae were highly resistant to Compound sulfamethoxazole and Ampicillin. Galanz negative bacteria had the lowest resistance to Piperacillin/ Tazobactam and Amikacin. Conclusions The main pathogens of CAP in children were G -bacteria. There were some differences among the isolates at different ages of CAP. Their resistance to very common anti-biotics was very high in children.