Animal biosafety laboratories generally use negative-pressure cages to raise animals,to guarantee biosafety.Laboratory monbeys are much larger than the rodents,so they are mostly housed in non-airtight negative pressure animal isolation equipments within animal biosafety laboratories.Regarding practical work experience in an Animal Biosafety Level 2(ABSL-2)laboratory,this study discusses how to control biological risks at various stages of monkey breeding and infectious experiments within an ABSL-2 laboratory.This exploration provides valuable insights and references for controlling biological risks in ABSL-2 and ABSL-3 laboratories carrying out infection experiments in monkeys in the future.

Objective To explore the effect of galangin(GAL)on bone metabolism by regulating Hippo/Yes associated protein(YAP)signaling pathway in rats with diabetic osteoporosis(DOP).Methods 90 female SD rats were randomly divided into normal control(NC)group,model(Mod)group,low-dose GAL(GAL-L)group,high-dose GAL(GAL-H)group,and high-dose GAL+YAP inhibitor Verteporfin(GAL-H+Verteporfin)group,with 18 rats in each group.Dual energy X-ray was used to detect bone mineral density(BMD).Three-point bending assay was used to evaluate the biomechanics of femur.HE staining was used to test the pathological changes in femoral tissue.ELISA was used to detect serum levels of osteocalcin(OC),alkaline phosphatase(ALP),and tartaric-resistant acid phosphatase(TRAP).Western blot was used to evaluate the expression of YAP protein in femoral tissue.Results Compared with NC group,the BMD,maximum load,elastic modulus,OC,ALP,and YAP protein expression of femur were reduced(P<0.05),and the TRAP was increased in Mod group(P<0.05).Compared with Mod group,BMD,maximum load,elastic modulus,OC,ALP and YAP protein expressions of femur were increased(P<0.05),while TRAP was decreased in GAL-L and GAL-H groups(P<0.05),and the improvement was better in GAL-H group.Compared with GAL-H group,femur BMD,maximum load,elastic modulus,OC,ALP and YAP protein expressions of femur were decreased(P<0.05),while TRAP was increased in GAL-H+Verteporfin group(P<0.05).Conclusions GAL may regulate the Hippo/YAP signaling pathway,increase the expression of YAP,regulate bone metabolism in DOP rats,and inhibit bone resorption.

Objective To investigate the effect of platelet lysate on mitochondrial function of astrocytes after spinal cord injury.Methods Astrocytoma cells SVGp12 were incubated with different concentrations of H2O2 (100 μmol/L,400 μmol/L,1000 μmol/L,2000 μmol/L),and the OD value was determined by CCK-8 assay. The H2O2 concentration with a cell inhibition rate of about 50％ was selected for follow-up experiments. The cells were incubated with different concentrations of platelet lysate (the volume ratios of platelet lysate to culture medium were 1∶10,1∶40,1∶80,1∶160 and 1∶320) for 24 hours,and the OD value was determined,thereby selecting the optimal concentration of platelet lysate based on the cell survival rate. SVGp12 cells were divided into the normal group (cultured normally without special treat-ment),the model group (incubated with 1000 μmol/L of H2O2 for 3 hours),and the treatment group (incubated with 1000 μmol/L of H2O2 for 3 hours and then treated with platelet lysate at a volume ratio of 1∶80). The mitochondrial function was evaluated by mitochondrial activity staining,mitochondrial membrane potential detection,and mitochondrial permeability transition pore detection. Results After incubating cells with 1000 μmol/L of H2O2 for 3 hours,the cell inhibition rate was 48％,and the OD value decreased significantly (P<0.01),confirming the successful establishment of a model of astrocytes with spinal cord injury. After treated with different concentrations of platelet lysate,the OD values and cell survival rate of the cells were higher than those in the model group (P<0.05). The concentration of platelet lysate (volume ratio of 1∶80) with cell viability of 20％ was selected for follow-up experiments. Platelet lysate could improve the morphology of injured astrocytes. The mitochondrial activity of the treatment group was significantly higher than that of the model group,and the mitochondrial activity of the model group was significantly lower than that of the normal group,with statistically significant differences (P<0.05). The mitochondrial membrane potential in the model group was lower than that in the normal group,and the mitochondrial membrane potential in the treatment group was higher than that in the model group,with statistically significant differences (P<0.01). The mitochondrial permeability in the model group was greater than that in the normal group,and the mitochondrial permeability in the treatment group was smaller than that in the model group,with statistically significant differences (P<0.01).Conclusion Platelet lysate can improve the survival rate of astrocytes after spinal cord injury,enhance the mitochondrial activity of cells,and improve the mitochondrial membrane potential and mitochondrial permeability transition pore opening.

Objective To investigate the relationship between prolactin and mild cognitive impairment(MCI)in postmenopausal women with type 2 diabetes mellitus(T2DM).Methods A total of 319 postmenopausal women with T2DM who were hospitalized in the Department of Endocrinology,Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University were enrolled in this study from August 2016 to October 2023.All the patients were divided into two groups according whether they had MCI:T2DM group(n=161)and MCI group(n=158).Differences in clinical characteristics were compared between the two groups.Pearson correlation was used to analyze the correlation between sex hormones and cognitive domains,and Logistic regression analysis was used to evaluate the influencing factors for MCI development.Results Serum prolactin levels were significantly lower in the MCI group than in the T2DM group[(5.5±2.1)vs(7.2±2.9)μg/L,P<0.05].Serum prolactin level was positively correlated with mini-mental state examination score,Montreal cognitive assessment score,immediate memory score,visuopatial constructional score,attention score and hippocampal volume(P<0.05),and negatively correlated with processing speed test(time)and executive function test(time)(P<0.05).Logistic regression analysis demonstrated that serum prolactin level was an influencing factor for the risk of MCI in postmenopausal women with T2DM(OR 0.715,95%CI 0.605～0.845,P<0.01).Conclusions The decrease of serum prolactin level is associated with an increased risk of MCI in postmenopausal women with T2DM.

Animal biosafety laboratories generally use negative-pressure cages to raise animals,to guarantee biosafety.Laboratory monbeys are much larger than the rodents,so they are mostly housed in non-airtight negative pressure animal isolation equipments within animal biosafety laboratories.Regarding practical work experience in an Animal Biosafety Level 2(ABSL-2)laboratory,this study discusses how to control biological risks at various stages of monkey breeding and infectious experiments within an ABSL-2 laboratory.This exploration provides valuable insights and references for controlling biological risks in ABSL-2 and ABSL-3 laboratories carrying out infection experiments in monkeys in the future.

Objective:To compare the safety and efficacy of 3D-printed porous bioceramic artificial bone and artificial bone substitutes in the treatment of limb bone defects.Methods:A total of 220 patients with post-traumatic limb bone defects admitted to Xijing Hospital Affiliated to Air Force Medical University of the Chinese People's Liberation Army (34 cases), the Third Hospital of Hebei Medical University (60 cases), Xi'an Honghui Hospital (28 cases), the Third Hospital of Southern Medical University (18 cases), Changsha Third Hospital (28 cases), Foshan Traditional Chinese Medicine Hospital (16 cases), Foshan Fuxing Chancheng Hospital (12 cases), and Henan Provincial Orthopaedic Hospital (24 cases) from May 2022 to October 2023 were included as research subjects. According to the manufacturing method of the bone graft material, the subjects were randomly divided into the 3D printed porous bioceramic artificial bone group (3D printing group) and the artificial bone substitute group (non-3D printing group) at a ratio of 1:1 by the envelope method. Adverse events that might be related to the surgery were selected through correlation evaluation and classified as abnormal laboratory indicators, systemic or other site symptoms and abnormalities, and local symptoms and abnormalities of the affected limb. The safety of the two groups was compared. The bone graft fusion rate, bone defect repair and healing rate, and short form 12 (SF-12) score of the two groups were calculated to evaluate the postoperative recovery.Results:Thirty-two cases were excluded (4 cases refused to use their data after reconsideration, 7 cases were not used after preoperative assessment, and 21 cases exceeded the standard for body mass index and laboratory indicators upon re-examination). A total of 188 cases were randomly divided into the 3D printing group and the non-3D printing group according to the random method, with 94 cases in each group. Among them, 11 cases in the 3D printing group and 9 cases in the non-3D printing group dropped out due to loss to follow-up. Finally, 168 cases completed the follow-up, including 83 cases in the 3D printing group and 85 cases in the non-3D printing group. In the 3D printing group, there were 53 males and 30 females, with an average age of 47.9±12.7 years; in the non-3D printing group, there were 53 males and 32 females, with an average age of 48.6±12.9 years. A total of 51 cases in the two groups experienced adverse events related to the surgery, including 13 cases of abnormal laboratory indicators (5 cases in the 3D printing group and 8 cases in the non-3D printing group), 15 cases of systemic or other site symptoms and abnormalities (9 cases in the 3D printing group and 6 cases in the non-3D printing group), and 23 cases of local symptoms and abnormalities of the affected limb (13 cases in the 3D printing group and 10 cases in the non-3D printing group). There was no statistically significant difference in the incidence of adverse events between the two groups ( P>0.05). The bone graft fusion rates of the 3D printing group and the non-3D printing group at 6 months after surgery were 99%(82/83) and 99%(84/85), respectively, and the bone defect repair and healing rates were 89%(74/83) and 89%(76/85), respectively. At the time of 12 months after surgery, the bone graft fusion rates were 99%(82/83) and 99%(84/85), respectively, and the bone defect repair and healing rates were 94%(78/83) and 92%(78/85), respectively. There was no statistically significant difference in the bone graft fusion rate and bone defect repair and healing rate between the two groups ( P>0.05). The SF-12 scores during the screening period were 27.82±2.96 points and 27.22±4.23 points in the 3D printing group and the non-3D printing group, respectively, and at 3 months after surgery were 28.08±3.13 points and 27.64±3.16 points, at 6 months after surgery were 29.42±3.10 points and 28.55±3.45 points, and at 12 months after surgery were 29.78±2.80 points and 29.58±2.94 points, respectively. There was no statistically significant difference between the groups ( P>0.05). Both groups of surgeries were successfully completed without any serious surgical or bone graft-related complications. Conclusion:The safety and efficacy of 3D-printed porous bioceramic artificial bone in the treatment of limb bone defects are not significantly different from those of currently clinically applied artificial bone substitutes.

Objective To investigate the relationship between prolactin and mild cognitive impairment(MCI)in postmenopausal women with type 2 diabetes mellitus(T2DM).Methods A total of 319 postmenopausal women with T2DM who were hospitalized in the Department of Endocrinology,Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University were enrolled in this study from August 2016 to October 2023.All the patients were divided into two groups according whether they had MCI:T2DM group(n=161)and MCI group(n=158).Differences in clinical characteristics were compared between the two groups.Pearson correlation was used to analyze the correlation between sex hormones and cognitive domains,and Logistic regression analysis was used to evaluate the influencing factors for MCI development.Results Serum prolactin levels were significantly lower in the MCI group than in the T2DM group[(5.5±2.1)vs(7.2±2.9)μg/L,P<0.05].Serum prolactin level was positively correlated with mini-mental state examination score,Montreal cognitive assessment score,immediate memory score,visuopatial constructional score,attention score and hippocampal volume(P<0.05),and negatively correlated with processing speed test(time)and executive function test(time)(P<0.05).Logistic regression analysis demonstrated that serum prolactin level was an influencing factor for the risk of MCI in postmenopausal women with T2DM(OR 0.715,95%CI 0.605～0.845,P<0.01).Conclusions The decrease of serum prolactin level is associated with an increased risk of MCI in postmenopausal women with T2DM.

Objective To explore the impact of different delay time of umbilical cord ligation on the outcome of preterm infants.Methods A total of 266 preterm infants born in the obstetric department of this hospital from January 2021 to December 2022 were selected as the research subjects and divided into the in-stant group(umbilical cord ligation immediately after birth,n=53),30 s group(delayed umbilical cord liga-tion for 30 s,n=50),60 s group(delayed umbilical cord ligation for 60 s,n=55),90 s group(delayed umbili-cal cord ligation for 90 s,n=55)and 120 s group(delayed umbilical cord ligation for 120 s,n=52)according to the random number table method.The Hb and HCT levels and the incidence rates of anemia,blood transfu-sion,hyperbilirubinemia,intracranial hemorrhage,necrotizing enterocolitis(NEC)and bronchopulmonary dys-plasia(BPD)after birth were compared among 5 groups.Results The Hb and HCT levels at 24 h after birth in the 60,90,120 s groups were higher than those in the instant group and 30 s group,moreover the 120 s group was higher than the 60 s group and 90 s group,and the differences were statistically significant(P＜0.05).The anemia incidence rate and blood transfusion rate in the 60,90,120 s groups were lower than those in the instant group and 30 s group,and the differences were statistically significant(P＜0.05).The hyperbil-irubinemia incidence rate in the 120 s group was higher than that in the other 4 groups,and the difference was statistically significant(P＜0.05).The intracranial hemorrhage incidence rate in the 60,90,120 s groups were lower than those in the instant group and 30 s group,and the differences were statistically significant(P＜0.05).The NEC incidence rate in the 60 s and 90 s group was lower than that in the instant group and 30 s group,but the 120 s group was higher than that in the other 4 groups,and the differences were statistically significant(P＜0.05).The BPD incidence rate had no statistical difference among 5 groups(P＞0.05).Con-clusion Delayed umbilical cord ligation for 60 s or 90 s could improve the outcomes of preterm infants.

Objective To explore the effect of galangin(GAL)on bone metabolism by regulating Hippo/Yes associated protein(YAP)signaling pathway in rats with diabetic osteoporosis(DOP).Methods 90 female SD rats were randomly divided into normal control(NC)group,model(Mod)group,low-dose GAL(GAL-L)group,high-dose GAL(GAL-H)group,and high-dose GAL+YAP inhibitor Verteporfin(GAL-H+Verteporfin)group,with 18 rats in each group.Dual energy X-ray was used to detect bone mineral density(BMD).Three-point bending assay was used to evaluate the biomechanics of femur.HE staining was used to test the pathological changes in femoral tissue.ELISA was used to detect serum levels of osteocalcin(OC),alkaline phosphatase(ALP),and tartaric-resistant acid phosphatase(TRAP).Western blot was used to evaluate the expression of YAP protein in femoral tissue.Results Compared with NC group,the BMD,maximum load,elastic modulus,OC,ALP,and YAP protein expression of femur were reduced(P<0.05),and the TRAP was increased in Mod group(P<0.05).Compared with Mod group,BMD,maximum load,elastic modulus,OC,ALP and YAP protein expressions of femur were increased(P<0.05),while TRAP was decreased in GAL-L and GAL-H groups(P<0.05),and the improvement was better in GAL-H group.Compared with GAL-H group,femur BMD,maximum load,elastic modulus,OC,ALP and YAP protein expressions of femur were decreased(P<0.05),while TRAP was increased in GAL-H+Verteporfin group(P<0.05).Conclusions GAL may regulate the Hippo/YAP signaling pathway,increase the expression of YAP,regulate bone metabolism in DOP rats,and inhibit bone resorption.

ObjectiveThis study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes. MethodsStructural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments. ResultsThe collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments. ConclusionThe antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.