BACKGROUND:Psoralen has a strong anti-osteoporotic activity and may have a restorative effect on chemotherapy-induced osteoporosis. OBJECTIVE:To explore the restorative effect of psoralen on the osteogenic differentiation of bone marrow mesenchymal stem cells in mice inhibited by cyclophosphamide and its mechanism. METHODS:C57BL/6 mouse bone marrow mesenchymal stem cells were isolated and cultured.Effect of psoralen on viability of bone marrow mesenchymal stem cells was detected by MTT assay.Osteogenic induction combined with alkaline phosphatase staining was used to determine the optimal dose of psoralen to restore the osteogenic differentiation of bone marrow mesenchymal stem cells inhibited by cyclophosphamide.The mRNA expression levels of Runx2,alkaline phosphatase,Osteocalcin,osteoprotegerin,and Wnt/β-catenin signaling pathway-related genes Wnt1,Wnt4,Wnt10b,β-catenin,and c-MYC were measured by RT-qPCR at different time points under the intervention with psoralen.The protein expression of osteogenic specific transcription factor Runx2 and Wnt/β-catenin signaling pathway related genes Active β-catenin,DKK1,c-MYC,and Cyclin D1 was determined by western blot assay at different time points under the intervention with psoralen. RESULTS AND CONCLUSION:(1)There was no significant effect of different concentrations of psoralen on the viability of bone marrow mesenchymal stem cells.The best recovery of the inhibition of osteogenic differentiation of bone marrow mesenchymal stem cells caused by cyclophosphamide was under the intervention of psoralen at a concentration of 200 μmol/L.(2)Psoralen reversed the reduction in osteogenic differentiation marker genes Runx2,alkaline phosphatase,Osteocalcin and osteoprotegerin mRNA expression and Runx2 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(3)Psoralen reversed the decrease in Wnt/β-catenin pathway-related genes Wnt4,β-catenin,c-MYC mRNA and Active β-catenin,c-MYC,and Cyclin D1 protein expression and the increase in DKK1 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(4)The results showed that cyclophosphamide inhibited osteogenic differentiation of bone marrow mesenchymal stem cells in mice,and psoralen had a restorative effect on it.The best intervention effect was achieved at a concentration of 200 μmol/L psoralen,and this protective effect might be related to the activation of Wnt4/β-catenin signaling pathway by psoralen.

BACKGROUND:Flap transplantation technique is a commonly used surgical procedure for the treatment of severe tissue defects,but postoperative flap necrosis is easily triggered by ischemia-reperfusion injury.Therefore,it is still an important research topic to improve the survival rate of transplanted flaps. OBJECTIVE:To review the pathogenesis and latest treatment progress of flap ischemia-reperfusion injury. METHODS:CNKI,WanFang Database and PubMed database were searched for relevant literature published from 2014 to 2024.The search terms used were"flap,ischemia-reperfusion injury,inflammatory response,oxidative stress,Ca2+overload,apoptosis,mesenchymal stem cells,platelet-rich plasma,signaling pathways,shock wave,pretreatment"in Chinese and English.After elimination of irrelevant literature,poor quality and obsolete literature,77 documents were finally included for review. RESULTS AND CONCLUSION:Flap ischemia/reperfusion injury may be related to pathological factors such as inflammatory response,oxidative stress response,Ca2+overload,and apoptosis,which can cause apoptosis of vascular endothelial cells,vascular damage and microcirculation disorders in the flap,and eventually lead to flap necrosis.Studies have found that mesenchymal stem cell transplantation,platelet-rich plasma,signaling pathway modulators,shock waves,and pretreatment can alleviate flap ischemia/reperfusion injuries from different aspects and to varying degrees,and reduce the necrosis rate and necrosis area of the grafted flap.Although there are many therapeutic methods for skin flap ischemia/reperfusion injury,a unified and effective therapeutic method has not yet been developed in the clinic,and the advantages and disadvantages of various therapeutic methods have not yet been compared.Most of the studies remain in the stage of animal experiments,rarely involving clinical observations.Therefore,a lot of research is required in the future to gradually move from animal experiments to the clinic in order to better serve the clinic.

OBJECTIVE To investigate the levels of procalcitonin(PCT),platelet-to-lymphocyte ratio(PLR),neu-trophil-to-lymphocyte ratio(NLR),white blood cell count(WBC)and serum amyloid A(SAA)in newborns with bloodstream infection,and their diagnostic value for early diagnosis of bloodstream infection in newborns.METHODS A total of 100 neonates with positive blood culture results from Jan.2021 to Jan.2024 at Taizhou Hos-pital of Zhejiang Province were selected.These cases were divided into three groups based on the type of pathogen:74 cases in the gram-negative bacterial group,19 cases in the gram-positive bacterial group,and 7 cases in the fungal group.Additionally,100 neonates with negative blood culture results during the same period were selected as the control group.By comparing the levels of PCT,PLR,NLR,WBC and SAA in the blood of newborns infec-ted with different pathogens,the diagnostic value of various indicators on the receiver operating characteristic(ROC)curve for diagnosis of bloodstream infection in newborns was evaluated.RESULTS Among 100 newborns with positive blood cultures,74 strains of gram-negative bacteria,19 strains of gram-positive bacteria,and 7 strains of fungi were identified.The levels of PCT,PLR,NLR,WBC and SAA in the gram-positive,gram-negative and fungal groups were all higher than those in the control group(P＜0.05).The ROC curve re-sults showed that the diagnostic efficacy of the combination of PCT,PLR,NLR,WBC and SAA for gram-nega-tive and gram-positive bloodstream infections was superior to that of each individual indicator(P＜0.05),with ar-ea under the curve(AUC)of 0.982 and 0.969,respectively.CONCLUSION PCT,PLR,NLR,WBC and SAA can serve as effective biomarkers for the early diagnosis of bloodstream infection in newborns,aiding in the rapid and accurate identification of the type of infection.

Objective:To explore the values of reticulin fiber staining (RFS) in evaluating bone marrow (BM) involvement of lymphoma and in grading of BM biopsy from adult lymphoma patients.Methods:Retrospectively,354 cases of adult lymphoma were collected from November 2023 to May 2024 at Peking University Cancer Hospital. BM samples were stained with RFS and immunohistochemical staining (IHC), and flow cytometry (FCM) was also performed with the BM aspirations simultaneously. RFS was graded according to the European Consensus, as high grade (grade 2-3) indicating BM involvement in the study. BM involvement was considered as definite if no less than two positive findings among IHC, FCM, and RFS. Statistical analyses were performed via SPSS software (V23.0).Results:In this series, 52.3% (185/354) of the patients were male; 35.0% (124/354) aged >60 years; BM involvements were found in 34.5% (122/354) cases with high grade of RFS, which, in turn, were lymphoblastic leukemia/lymphoma (ALL/LBL) group (4/4), indolent B-cell lymphoma (IndBCL) group (49.1%, 53/108), transformed B-cell lymphoma (TrBCL) group (2/5), invasive B-cell lymphoma (InvBCL) group (26.5%, 41/155), T and NK cell lymphoma (TNKCL) group (27.3%, 12/44) and classical Hodgkin lymphoma (CHL) group (26.3%, 10/38); if classified by specific types, T-ALL/LBL (2/2), B-ALL/LBL (2/2) and CLL/SLL (8/10) ranked top three. In terms of the positive rate of BM involvement evaluated by RFS, no significant difference was seen between either gender or age groups ( χ2=3.416, P=0.332 and χ2=4.200, P=0.241); however, significant differences were observed between different lymphoma groups and types ( χ2=29.961, P=0.012 and χ2=102.546, P<0.001, respectively). BM invasion rates indicated by IHC and FCM were 25.4% (90/354) and 13.8% (49/354), respectively. The overall BM invasion rate was 24.3% (86/354), and the sensitivity of RFS, IHC, and FCM was 90.8%, 97.8%, and 55.8%, and specificity was 84.1%, 99.6%, and 98.9%, respectively. Overall, the concordance rate of RFS with IHC and FCM was 83.6% and 74.0%, respectively, including 85.8% and 74.2% for InvBCL group, 79.6% and 75.0% for IndBCL group, 84.1% and 75.0% for TNKCL group, 81.6% and 73.7% for CHL group, 5/5 and 2/5 for TrBCL group, and 4/4 and 3/4 for ALL/LBL group. Conclusions:In the evaluation of BM involvement status of adult lymphoma, high sensitivity and specificity are observed by RFS, and high concordance is also noted with both IHC and FCM. Thus, the BM infiltrating status of adult lymphoma could be evaluated more accurately by a combined usage of the three methods.

Purpose To investigate the predictive value of the methylation regulator heterogeneous nuclear ribonu-cleoprotein C(HNRNPC)in the extracellular vesicles(EVs)derived from cervical cancer cells for tumor invasion and metastasis.Methods In vitro experiments were conducted using a human normal cervical epithelial cell line(Hcer-Epic),three cervical cancer cell lines(HeLa,SiHa and CaSki)and human umbilical vein endothelial cells(HU-VECs).HNRNPC protein expression was assessed by Western blot.CaSki cells were transfected with either sh-NC or sh-HNRNPC,and EVs were subsequently isolated from culture supernatant.The effects of EVs on the proliferation,migration and invasion of SiHa cells,as well as on angiogenesis in HUVECs,were investigated respectively.For tissue microarray analysis,normal cervical tissues(n=8),low-grade squamous intraepithelial lesions(LSIL,n=32)and high-grade squamous intraepithelial lesions(HSIL,n=37),and cervical carcinoma specimens(n=153)were ob-tained from our institution.Based on HNRNPC immunoscores,cervical cancer patients were divided into two groups:high(n=99)and low(n=54)HNRNPC expression group,and their clinicopathological features and prognosis were compared.Results Compared with HcerEpic cells,the expression of HNRNPC increased in SiHa,HeLa and CaSki cells.In SiHa cells treated with EVs from CaSki cell,the group receiving EVs from sh-HNRNPC-transfected CaSki cells showed significantly reduced proliferation,colony formation,migration,and invasion compared with the sh-NC-EVs group(P＜0.05).Similarly,in HUVECs treated with CaSki-EVs,compared with sh-NC-EVs group,the sh-HNRNPC-EVs group demonsteated significantly lower protein expression of PCNA and VEGFA and reduced angiogene-sis length(P＜0.05).HNRNPC expression gradually increased during the transformation of cervical epithelial cells(F=106.9,P＜0.001),and was significantlyelevated in HSIL and cervical cancer tissues compared with normal tis-sues(P＜0.001).Moreover,in cervical cancer,high HNRNPC expression was significantly related to poorer cellular differentiation,larger tumor size,parametrial and vaginal infiltration,advanced FIGO stage,and pelvic lymph node metastasis(P＜0.05).Kaplan-Meier analysis showed that that patients with high HNRNPC expression had significant-ly shorter overall survival post-surgery compared with those with low expression(P＜0.05).Conclusion EVs contai-ning HNRNPC contribute to cervical cancer progression by promoting tumor growth,invasion,and angiogenesis,there-by accelerating metastasis.In addition,high HNRNPC expression in cervical cancer tissue is related to poor prognosis of patients,indicating that EV-associated HNRNPC may serve as a potential therapeutic target.

Objective To find out whether photobiomodulation(PBM)can mitigate cognitive dysfunction caused by chronic stress by affecting levels of adenosine triphosphate(ATP)and adenosine receptors.Methods Twenty-four C57BL/6J mice were randomly divided into a control group,a stress group,and a treatment group.Chronic unpredictable mild stress was used to establish a mouse model of stress.Six weeks into modeling,the treatment group was subjected to one week of PBM interventions.Behavioral tests were conducted to observe behavioral changes in the mice.Western blotting(WB)was used to detect the expressions of A1,A2B,and A3 adenosine receptors in the hippocampus and prefrontal cortex of mice in the three groups.Twelve C57BL/6J mice were randomly divided into a control group and an intervention group.The intervention group received a week of PBM interventions and underwent behavioral testing.WB was used to detect the expression changes of A1,A2B,and A3 adenosine receptors in the hippocampus and prefrontal cortex in both groups.Immunofluorescence assay was adopted to detect the expression of c-Fos in the hippocampus of mice in the two groups.The ATP assay kit made by Beyotime Biotechnology Co.,Ltd.was used to measure changes in ATP contents in the hippocampus and prefrontal cortex tissues of mice.Cell experiments were conducted to verify the effect of PBM on intracellular ATP contents.Results Mice in the stress group covered a similar distance to the control group,but finished far fewer platform crossings.There was no significant difference between the treatment group and the control group in the number of times of platform crossings,but compared favorably with the stress group where the levels of adenosine receptors in the hippocampus and prefrontal cortex were lower,but were increased by PBM.After PBM interventions in normal mice,platform crossings were increased significantly compared to the control group.PBM also raised adenosine receptor levels and ATP contents in the hippocampus and prefrontal cortex,and increased hippocampal c-Fos expressions.In vitro,PBM elevated intracellular ATP levels.Conclusion PBM may improve chronic stress-induced cognitive dysfunction by regulating ATP levels and adenosine receptor expressions,thereby modulating neuronal responsiveness in the hippocampus.

Objective:To explore the values of reticulin fiber staining (RFS) in evaluating bone marrow (BM) involvement of lymphoma and in grading of BM biopsy from adult lymphoma patients.Methods:Retrospectively,354 cases of adult lymphoma were collected from November 2023 to May 2024 at Peking University Cancer Hospital. BM samples were stained with RFS and immunohistochemical staining (IHC), and flow cytometry (FCM) was also performed with the BM aspirations simultaneously. RFS was graded according to the European Consensus, as high grade (grade 2-3) indicating BM involvement in the study. BM involvement was considered as definite if no less than two positive findings among IHC, FCM, and RFS. Statistical analyses were performed via SPSS software (V23.0).Results:In this series, 52.3% (185/354) of the patients were male; 35.0% (124/354) aged >60 years; BM involvements were found in 34.5% (122/354) cases with high grade of RFS, which, in turn, were lymphoblastic leukemia/lymphoma (ALL/LBL) group (4/4), indolent B-cell lymphoma (IndBCL) group (49.1%, 53/108), transformed B-cell lymphoma (TrBCL) group (2/5), invasive B-cell lymphoma (InvBCL) group (26.5%, 41/155), T and NK cell lymphoma (TNKCL) group (27.3%, 12/44) and classical Hodgkin lymphoma (CHL) group (26.3%, 10/38); if classified by specific types, T-ALL/LBL (2/2), B-ALL/LBL (2/2) and CLL/SLL (8/10) ranked top three. In terms of the positive rate of BM involvement evaluated by RFS, no significant difference was seen between either gender or age groups ( χ2=3.416, P=0.332 and χ2=4.200, P=0.241); however, significant differences were observed between different lymphoma groups and types ( χ2=29.961, P=0.012 and χ2=102.546, P<0.001, respectively). BM invasion rates indicated by IHC and FCM were 25.4% (90/354) and 13.8% (49/354), respectively. The overall BM invasion rate was 24.3% (86/354), and the sensitivity of RFS, IHC, and FCM was 90.8%, 97.8%, and 55.8%, and specificity was 84.1%, 99.6%, and 98.9%, respectively. Overall, the concordance rate of RFS with IHC and FCM was 83.6% and 74.0%, respectively, including 85.8% and 74.2% for InvBCL group, 79.6% and 75.0% for IndBCL group, 84.1% and 75.0% for TNKCL group, 81.6% and 73.7% for CHL group, 5/5 and 2/5 for TrBCL group, and 4/4 and 3/4 for ALL/LBL group. Conclusions:In the evaluation of BM involvement status of adult lymphoma, high sensitivity and specificity are observed by RFS, and high concordance is also noted with both IHC and FCM. Thus, the BM infiltrating status of adult lymphoma could be evaluated more accurately by a combined usage of the three methods.

Objective To establish a dexamethasone(Dex)-induced zebrafish model of glucocorticoid-induced osteoporosis(GIOP)combined with glucocorticoid-induced hypertension(GIHT),and to validate the model by the systematic evaluation of both the phenotypic manifestations and molecular mechanisms.Methods Zebrafish larvae at 3 or 4 d post-fertilization(dpf)were divided randomly into a control group(0.1％dimethyl sulfoxide)and a model group(10 μmol/L Dex).Osteogenic parameters and vessel diameter were assessed at 0,48,and 96 h post-administration(n=10).Bone mineralization and density were determined by the total area and sum brightness after Alizarin red staining.Vessel diameter was measured by detecting blood flow in the dorsal aorta.After confirming the optimal administration time,expression levels of bone-formation-related proteins(protein kinase B(Akt),glycogen synthase kinase(GSK)-3β,β-catenin)and angiogenesis-related proteins(AMP-activated protein kinase(AMPK),nuclear factor(NF)-κB)were detected by Western Blot to verify the molecular effectiveness of the model.Results Exposure to Dex for 96 h reduced bone mineralization and density in zebrafish larvae compared with the control group,and statistical analysis identified 4 dpf zebrafish and Dex administration for 96 h as the optimal modeling times for the GIOP model.Blood vessel diameter was significantly decreased in the model group compared with the control group(P＜0.05),and the difference became more pronounced with longer administration time and was particularly evident at 4 dpf and treatment for 96 h.Western Blot analysis showed that Dex significantly decreased protein expression levels of Akt,β-catenin,and NF-κB(P＜0.05)and significantly increased the expression of GSK-3β and AMPK(P＜0.05),suggesting that Dex effectively inhibited bone formation and angiogenesis after 96 hours treatment in 4 dpf zebrafish.Conclusions Treatment of 4 dpf zebrafish larvae with 10 μmol/L Dex rapidly established a reliable zebrafish model of GIOP combined with GIHT,providing an ideal animal model for further studies of the common mechanisms of the two diseases and for screening new drugs.

Objective:To explore the degree of change in anteversion angle and related risk factors after intramedullary nail fixation of intertrochanteric femur fracture in the elderly.Methods:The data of 256 elderly patients who underwent intramedullary nail fixation for intertrochanteric fractures of the femur at Tianjin Hospital of Tianjin University from March 2020 to March 2023 were selected, including 114 males and 142 females, with an age of 75.40±10.69 years (range, 65-94 years). The degree of change in the anteversion angle of the affected hip before and after the surgery was measured by CT scan of the hip, the Receiver Operating Characteristic Curve (ROC) was plotted, the area under the ROC curve was analyzed, and the optimal degree of grouping was determined by calculating the Youden Index, then all the patients were divided into two groups. The correlation between various risk factors (age, sex, type of internal fixation, fracture AO type, quality of reduction, fracture medial cortical defect or not, cusp distance) and the change of anterior tilt angle was screened by univariate and multivariate logistic regression analyses.Results:All 256 patients were followed up for 20.7±2.1 months (range, 18-23 months). Anteversion on the healthy side was 12.68°±5.10° (range, 5°-28°); postoperative anteversion on the affected side was 15.04°±7.67° (range, 9°-36°). By comparing the difference in the anterior tilt angle between the affected side and the healthy side, it was found that the anterior tilt angle of 67 patients was completely restored to the healthy side level after the operation. The anteversion angle was enlarged in 131 cases, of which the mildly increased angle (1°-9°) was found in 106 cases, moderately increased (10°-15°) was found in 17 cases, and significantly increased (>15°) was found in 8 cases; 58 patients showed anteversion angle reduction, of which 45 cases were mildly reduced (1°-9°), 13 cases were moderately reduced (10°-14°). The area under the ROC curve for the patient's anteversion angle and its 95% CI were 0.714(0.559, 0.867), and the maximum value of its Youden Index was 0.221, which corresponded to the optimal critical angle of 4°. There was no statistically significant difference in age, gender, reduction quality or fracture AO classification between the group with an anteversion angle>4° and the group with an anteversion angle≤4° ( P>0.05). The types of internal fixation, medial cortical defect and insufficient tip apex distance (TAD) were included in the binary variable logistic regression analysis. The results showed that single-nail internal fixation [ OR=0.412, 95% CI(0.244, 0.695), P=0.007], medial cortical defect [ OR=0.471, 95% CI(0.279, 0.793), P=0.009] and TAD>25 mm [ OR=0.367, 95% CI(0.207, 0.651), P=0.003] are independent risk factors for changes in anteversion angle after intramedullary nail fixation of intertrochanteric femur fractures in elderly. Conclusion:Single-nail internal fixation, medial cortical defect and TAD >25 mm are independent risk factors for the change of anteversion angle after intramedullary nail internal fixation of intertrochanteric fractures in the elderly.

Objective:To understand the clinical characteristics of myelopathy induced by intrathecal chemotherapy of methotrexate (MTX) and/or cytarabine (Ara-C).Methods:Relevant databases at home and abroad (up to February 18, 2024) were searched and case reports of myelopathy induced by intrathecal chemotherapy of MTX and/or Ara-C were collected. The patients′ general situation (gender, age, primary disease, etc.), use of MTX and/or Ara-C, previous radiotherapy, and occurrence time, clinical manifestations, spinal magnetic resonance imaging (MRI) results, cerebrospinal fluid test results, treatments and outcomes of myelopathy were extracted and analyzed descriptively and statistically.Results:A total of 75 articles were enrolled, involving 104 patients, with 62 males, 35 females, and 7 unknown genders. Their ages ranged from 1 to 74 years, with a median age of 26 years. The primary diseases included hematological malignancy in 101 cases, and other solid tumors in 3 cases. Before the occurrence of myelopathy, 42 cases had central nervous system tumor infiltration. Seventy-three patients received intrathecal injection of MTX combined with Ara-C, 21 patients received single MTX therapy, 10 patients received single Ara-C therapy. The number of intrathecal injections ranged from 1 to 62, with a median of 5 injections. Twenty-nine patients had received radiotherapy before. When myelopathy occurred, the cumulative dose of MTX was 7.5-480.0 mg, with a median cumulative dose of 60.0 mg; the cumulative dose of Ara-C was 15-1 599 mg, with a median cumulative dose of 280 mg. The onset time of myelopathy was from immediately to 365 days after the last intrathecal injection, with a median time of 2 days. The main clinical manifestations were weakness of both lower limbs, urinary and fecal incontinence or retention, paresthesia, and paraplegia, etc. Fifty-three patients had spinal abnormality in MRI examination, 32 had abnormal cerebrospinal fluid protein quantity, intrathecal basic protein, or homocysteine. After the diagnosis of myelopathy, 86 patients were treated with drugs, radiotherapy, plasma exchange, and cerebrospinal fluid exchange, and 18 patients had no record of treatment situation. Therapeutic agents included glucocorticoids, B vitamins, folic acid, immunoglobulin, leucovorin, S-adenosylmethionine, and dextromethorphan. Of the 104 patients, 20 achieved complete remission, with a median remission time of 30 hours; 25 experienced partial remission, with a median duration of 120 days; 32 showed no significant improvement; 26 died; one patient′s prognosis and outcome were unknown.Conclusions:The median occurrence time of myelopathy induced by intrathecal injection of MTX and/or Ara-C is 2 days. The main clinical manifestations are bilateral lower extremity weakness, urinary and bowel incontinence or retention, paresthesia, and paraplegia, etc. Abnormal spinal in MRI examination, quantitative cerebrospinal fluid protein, intrathecal basic protein occurred in some patients. Intrathecal injection should be stopped immediately after diagnosis of myelopathy, and the treatments such as drug and cerebrospinal fluid replacement should be given. The clinical outcome of myelopathy induced by intrathecal MTX and/or Ara-C was poor.