Objective:To construct a novel biomimetic calcium phosphate (BioCaP) scaffold loaded with bone morphogenetic protein-2 (BMP-2),and to investigate its role in the osteogenesis of human adipose-derived stem cells (hASCs) in vitro and in vivo.Methods:The BioCaP scaffold coprecipitated with BMP-2 (BMP-2-BioCaP) was constructed in this study.Field emission scanning electron microscopy (SEM) was used to analyze the morphology of the surfaces.The release kinetics was measured to evaluate the slow-release characteristics in vitro.BMP-2-BioCaP was immersed in proliferation medium (PM) or osteogenic medium (OM),respectively.The supernatants were collected and used to culture hASCs in vitro.Cell numbers were determined using the cell-counting kit-8 (CCK-8) to assess the cell proliferation.After 7 and 14 days,alkaline phosphatase (ALP) staining and quantification were performed to test the activity of ALP.After 14 and 21 days,the calcification deposition was determined by alizarin red S (ARS) staining and quantification.The expressions of the osteoblast-related genes were tested on day 4 and day 14.In the in vivo study,6 nude mice were used and implanted subcutaneously into the back of the nude mice for 4 groups:(1) BioCaP scaffold only,(2) BioCaP scaffold + hASCs,(3) BMP-2-BioCaP scaffold,(4) BMP-2-BioCaP scaffold + hASCs (test group).After 4 weeks of implantation,hematoxylin-eosin (HE) staining was performed to evaluate the in vivo osteogenesis of hASCs.Results:SEM observations showed that BioCaP and BMP-2-BioCaP scaffold were entirely composed of straight,plate-like and sharp-edged crystal units,and the length of the crystal units varied between 5 and 10 μm.Release kinetics analysis demonstrated that BMP-2 incorporated with BioCaP could be released at certain concentration and last for more than 21 days,and the accumulative protein release could reach 20％.CCK-8 assays showed that cell proliferation was not significantly affected by BMP-2BioCaP.ALP activity was higher by the induction of OM + BMP-2-BioCaP than of the other groups (P ＜0.01).More mineralization deposition and more expressions of osteoblast-related genes such as Runt-related transcription factor 2 (RUNX2),ALP,osteopontin (OPN) and osteocalcin (OC) were determined in the OM + BMP-2-BioCaP group at different time points (P ＜0.01).HE staining showed that,in the test group and BMP-2-BioCaP scaffold group,the extracellular matrix (ECM) with eosinophilic staining were observed around hASCs,and newly-formed bone-like tissues could be found in ECM around the scaffold materials.Moreover,compared with the BMP-2-BioCaP scaffold group,more bone-like tissues could be observed in ECM with typical structure of bone tissue in the test groups.No obvious positive results were found in the other groups.Conclusion:BMP-2-BioCaP scaffold could achieve slow-release of BMP-2 and promote the osteogenic differentiation of hASCs in vitro and in vivo.The novel tissue-engineered bone composed of hASCs and BMP-2-BioCaPis promising for the repair of bone defect.

Objective To elaborate the role of hepatic inherent macrophages and monocyte-derived macrophages in acute liver injury.Methods A model of acute liver injury in mice was induced via intraperitoneally injection of CCl 4 .Af-ter CCl4 injection, analysis of the expression of CD45, F4/80, Ly6C and CD11b on the hepatic macrophage surface was performed by flow cytometry at 24, 48 and 72 h before the hepatic inherent macrophages (CD45+F4/80hi) and monocyte-de-rived macrophages ( CD45+F4/80 lo ) were sorted at the same time .The relative expression of cytokines in the two popula-tions of macrophages was detected by qRT-PCR.Results Compared with control , the number of total F4/80 +cells in the liver was markedly increased after CCl 4 injection, especially at 72 h.The number of CD45+F4/80lo cells increased signifi-cantly after CCl4 injection 24 h.The mRNA levels of MCP-1, TNF-α, TGF-β1, matrix metalloproteinase(MMP)-12 and MMP-13 were elevated significantly both in F 4/80 hi and F4/80 lo macrophages .IL-12βand IL-6 mRNA levels increased significantly only in F4/80hi macrophages, while the level of MMP-9 mRNA increased markedly only in F4/80lo macropha-ges.When compared with F4/80lo macrophages, MCP-1 and MMP-12 mRNA levels were elevated significantly , while the level of TNF-αmRNA decreased significantly in F4/80hi macrophages.Conclusion In acute liver injury, hepatic inherent macrophages and monocyte-derived macrophages both express inflammatory cytokines , promoting inflammation response and leading to liver damage .The ability to recruit inflammatory monocytes into the liver is much stronger ,the expression of in-flammatory cytokines ( IL-12βand IL-6 ) and MMP-12 is higher , but the expression of inflammatory cytokines ( TNF-α) MMP-9 is lower in hepatic inherent macrophages than in monocyte-derived macrophages .

Objective To understand the clinical characteristics, treatment effect and prognosis of children with methylmalonic acidemia (MMA) combined with renal damage, and to provide experiences about how to improve the level of diagnosis and treatment.Methods The medical records of children with MMA were collected from January of 2007 to December of 2013 in Beijing Children's Hospital Affiliated to Capital University of Medical Sciences,while the clinical manifestations,laboratory findings, imaging material, inspection results were analyzed, and the renal pathological gene analysis, treatment effect, and prognosis of MMA children with renal damage were studied, and follow-up was carried out for 3 months up to 7 years.Results Among the two hundred and ninety-six cases of MMA,28 cases (9.5％) with renal damage,including 19 boys and 9 girls, 18 patients with hyperhomocysteinemia.Their ages varied from 2 days to 13 years old when confirmed.The first symptoms of 16 cases were proteinuria, hematuria, and edema, 9 cases were complicated with acute renal failure,5 cases were diagnosed as hemolytic uremic syndrome,6 patients were diagnosed as nephrotic syndrome, receiving enough glucocorticoids treatment without improvement.The routine examination of the urine showed that proteinuria (241-3 060 mg/d), methylmalonic acid levels and urinary protein were positively correlated (r =0.982, P =0.003), as urinary β2-microglobulin [(360.386 ± 231.496) μg/L], retinol binding protein [(338.537 ± 243.293) μg/L] increased.Extrarenal clinical manifestations were the spiritual, mental and physical development retardations, seizures, anemia etc.Two cases had a positive family history.Renal pathology showed mesangial cells and mesangial matrix proliferation broadening, no electron dense deposits in mesangial area, renal tubular epithelial cell swelling degeneration, and immunofluorescence was negative.Gene analysis of 3 cases showed exon1:c.80A ＞ G,p.Q27R and exon1 :c.365A ＞ T,p.H122L/c.609 G ＞ A,p.W203X.Children with MMA were treated with vitamin B12 treatment.Two cases of multiple organ failure died during hospitalization, and 2 cases died after leaving hospital.After treatment, the level of urinary methylmalonic acid was significantly decreased, while nervous system symptoms improved significantly, and the mental state and the renal function were improved.Conclusions MMA may be associated with kidney damage,usually with severe renal tubular and glomerular impairment.Early diagnosis is the key,and timely treatment can effectively control the disease, improve the prognosis.

Objective To analyze the clinical features and the correlation between radiographic parameters and incidence of calcaneal spur in the patients from Nanfang Hospital,Southern Medical University,China.Methods Three experienced observers independently used the image acquisition and transmission system (PACS) to collect the data of lateral and axial X-ray images of calcaneus or ankle joint in neutral position from the patients with calcaneal spur and normal controls who had undergone radiological examination in Nanfang Hospital,Southern Medical University from July 2014 through December 2015.Ten radiological parameters of the foot (B(o)hler angle,Gissane angle,calcaneal inclination angle,talocalcaneal angle,talus horizontal angle,posterior facet inclination angle,calcaneal length,height of the posterior facet,absolute foot height,and calcaneal width) were measured in both the patients and normal controls.The location,morphology and length of calcaneal spurs were compared between genders,sides and age groups.Results A total of 216 parpatients were included in the study.Female patients were more than male ones,simple plantar spurs more than simple achilles tendon ones,type B spurs more than type A ones,the length of achilles tendon spurs larger than that of plantar ones,female plantar spurs more than males ones,and the length of right foot plantar spurs larger than that of left foot ones.All the differences above were statistically significant (P ＜ 0.05).There were no significant differences in the location,morphology or length of calcaneal spurs between the age group of ≤ 60 years old and the age group of ＞ 60 years old (P ＞ 0.05).The incidence of calcaneal spur were significantly correlated to Gissane angle (P =0.000,OR =0.944,95％ CI 0.917-0.973),posterior facet inclination angle (P=0.017,OR=0.957,95％ CI 0.924-0.992) and height of the posterior facet (P =0.007,OR =0.933,95％ CI O.886-0.981).Conclusions Calcaneal spur favored more females than males.Plantar spurs were more common than Achilles ones.Plantar spurs of Type B were more common than those of Type A.Achilles spurs were longer than plantar ones.More females suffered plantar spur than males.Right foot spurs were longer than left foot ones.Age had no significant influence on the spur characteristics.The incidence of spur might have been related to the Gissane angle,posterior facet inclination angle and height of the posterior facet of the foot.

Objective:To explore the diagnostic value of the toxicant and drug detection in clinical poisoning diseases and analyze the clinical characteristics of patients with positive poison test.Methods:This study was a multicenter retrospective cohort study. Sampling and clinical information data were collected between October 1, 2020 and September 30, 2022 from 41 tertiary hospitals in and around Jiangsu province. The clinical characteristics of patients with positive toxicology tests were analyzed, and the correlation between the drug sampling situation and the test results was analyzed..Results:A total of 895 patients with clinical diagnosis or suspected poisoning were enrolled in this study. Among them, 652 patients had positive results, accounting for 72.85%. Among all positive patients, 506 patients were exposed to a single poison and 147 patients were exposed to multiple poisons. The top three poisons were pesticide herbicides (202 cases, 30.98%), sedative and psychotropic drugs (151 cases, 23.16%), and pesticide insecticides (97 cases, 14.88%). Among 541 patients with clear exposure history, the positive rate was 78.19%, and among 354 patients with unclear exposure history, the positive rate was 64.69%. The top three poisons (drugs) of patients with unclear exposure history were sedative and psychotropic (82, 12.58%), herbicide (26, 3.99%), and rodenticide (22, 3.37%). Patients who admitted to hospital for unexplained consciousness disorder, abnormal blood coagulation function and multiple organ dysfunction were more likely to obtain positive poison test results.Conclusions:There is uncertainty in the exposure history of poisoning diseases, so it is necessary to improve the detection of toxic substances as soon as possible. Toxicant testing should be considered when patients have impaired consciousness, abnormal coagulation function and multiple organ dysfunction.

Objective:To explore the clinical characteristics of poisoned patients with poisons purchase online.Methods:A retrospective case-control study was conducted on poisoned patients purchased poisons online from 1st January 2021 to 31th May 2022 in the Emergency Department of the First Affiliated Hospital of Nanjing Medical University. The clinical data including sex, age, way of medical treatment, cause of poisoning, exposure routes, category of toxic drugs, gastric lavage, toxic detection and prognosis of patients were collected and compared with those patients obtained poisons at stores as the control group.Results:Totally 318 poisoned patients were included in this study, of which 44 (13.8%) were obtained poisons online. Compared with the patients obtained poisons at stores, the patients obtained poisons online were younger ( P<0.001), and had higher proportion of suicide intention ( P=0.006), more oral route exposure ( P=0.029), and more proportions of receiving gastric lavage before transfer to the hospital ( P=0.001). Pesticides and fertilizers with organic heterocycles were the main types of poisons in the online group, and there was no statistical difference in the distribution of poisons compared with the control group. Mixed drug poisoning was the leading cause in both online group (27.8%) and control group (38.8%) in drug overdose poisoned types, followed by dextromethorphan (16.7%) and estazolam (15.5%) in the online group. Conclusions:Young people are the main group getting poisons through the Internet. Health education should be strengthened for this group, and online shopping platforms should pay attention to the poisoning risk of potential overdose drugs or poisons transactions.

The clinical data of a case with late-onset isolated sulfite oxidase deficiency(ISOD)admitted in the Department of Neurology, Children′s Hospital, Zhejiang University School of Medicine in July 2021 were retrospectively analyzed.Fifteen previously published cases of late-onset ISOD were also reviewed.The patient was a girl, who was hospitalized because of " motor regression with mental retardation for 5 days" at 1 year old.The manifestations of the patient were extrapyramidal symptoms, regression of motor development and seizures.The level of urinary sulfites in the patient was increased.Magnetic resonance imaging (MRI) features were bilateral pallidus and substantia nigra.Gene sequencing suggested a pure missense mutation of the sulfite oxidase( SUOX) gene c. 650(exon5)G>A(p.Arg217Gln). In 16 cases of late-onset ISOD, the median age at onset and diagnosis was 10.5 months and 34.0 months, respectively.The common clinical manifestations were hypotonia (13 cases), seizures (10 cases), movement disorders (9 cases), and ectopia lentis (6 cases). The most common brain MRI feature was pallidus changes (11 cases), followed by lesions of substantia nigra (5 cases), and cerebral atrophy (4 cases). Fourteen cases of late-onset ISOD showed a positive urinary sulfite test.The missense mutation of the SUOX gene was found in 9 cases.It suggested that brain MRI involvement of bilateral pallidus, high excretion of urine sulfites and the missense mutation of the SUOX gene were important diagnostic clues for late-onset ISOD.

ObjectiveTo observe the effect of Danshen injection （DAN） on platelet （PLT）-induced metastasis of breast cancer cells in vitro. MethodThe 3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide （MTT） assay was used to observe the effect of DAN on the growth of MDA-MB-231 cells in vitro. Oris™ migration assay was used to determine the effect of DAN （final mass concentrations 4， 8， 16 g·L^-1） on PLT （1.5×10¹⁰ cells/L）-induced migration of breast cancer cells in vitro. The effect of DAN on PLT-induced cell invasion was detected by Transwell assay. Immunofluorescence and Western blot were used to detect the effect of DAN on the protein expression associated with PLT-induced epithelial-mesenchymal transition （EMT）. In addition， enzyme-linked immune-sorbent assay （ELISA） was used to determine the effect of DAN （final mass concentrations 4， 8， 16， 32， 64 g·L^-1） on the secretion of transforming growth factor-β₁ （TGF-β₁）. Western blot was used to observe the effect of DAN on the expression of podoplanin （PDPN） protein in MDA-MB-231 cells induced by PLT. ResultCompared with the blank group， the DAN groups （32 and 64 g·L^-1） showed decreased A₅₇₀ （P<0.05， P<0.01）， and there was no significant difference in A₅₇₀ between DAN groups （4， 8， 16 g·L^-1）. Compared with the blank group， the PLT group showed increased cell migration and invasion， while DAN groups significantly inhibited PLT-induced cell migration and invasion. Compared with the blank group， the PLT group showed decreased expression of E-cadherin， while DAN could significantly reverse this effect of PLT. Compared with the blank group， the PLT group showed increased Slug and Snail protein expression （P<0.05， P<0.01）， while DAN significantly reversed Snail protein expression induced by PLT （P<0.05， P<0.01）. The content of TGF-β₁ in the PLT group increased （P<0.01）， while the secretion of TGF-β₁ induced by PLT decreased in the DAN groups （16， 32， and 64 g·L^-1） （P<0.05， P<0.01）， and the secretion of TGF-β₁ was not significantly affected in other DAN groups. PDPN protein expression in the PLT group increased （P<0.01）， while DAN could significantly inhibit PLT-induced PDPN expression （P<0.01）. ConclusionDAN can inhibit PLT-induced migration， invasion， and EMT of breast cancer cells. The mechanism may be related to the direct action between breast cancer cells and tumor cells by down-regulating PDPN expression and interfering with PLT and has nothing to do with the effect of TGF-β₁ secretion of PLT.

ObjectiveTo observe the clinical efficacy of Chinese medicine combined with indirect moxibustion plaster on corona virus disease 2019 （COVID-19） patients during recovery period. MethodNinety patients of COVID-19 during the recovery period were randomly divided into a Chinese medicine group， an indirect moxibustion plaster group， and a combination group，with 30 cases in each group. According to the 10th edition of COVID-19 Diagnosis and Treatment Protocol，patients in the Chinese medicine group received oral Chinese medicine based on syndrome differentiation，one dose per day， twice a day. Patients in the indirect moxibustion plaster group were treated with indirect moxibustion plaster at Zusanli （ST 36）， Pishu （BL 20）， Dazhui （GV 14）， Feishu （BL 13）， Kongzui （LU 6）， and Tiantu （CV 22），once a day，40 min each time. Patients in the combination group were treated with Chinese medicine combined with indirect moxibustion plaster. Treatment lasted two weeks. Before and after treatment，the traditional Chinese medicine （TCM） symptom score，pulmonary computed tomography （CT） score，St. George's Respiratory Questionnaire （SGRQ） score，blood routine indexes ［white blood cell count （WBC），neutrophil count （NEUT），and lymphocyte count （LYM）］， and inflammatory indexes ［C-reactive protein （CRP），serum ferritin， and interleukin-6 （IL-6）］ were observed in the three groups. The clinical efficacy was evaluated. ResultAfter treatment，the scores of TCM symptoms，pulmonary CT， and SGRQ，CRP，IL-6，and ferritin in the three groups decreased（P<0.05），while WBC and LYM increased（P<0.05）， but there was no significant difference in NEUT. The above indexes in the combination group were better than those in the other two groups（P<0.05）. After treatment， the cured and markedly effective rate was 76.7% （23/30） in the combination group， 50.0% （15/30） in the Chinese medicine group， and 46.7% （14/30） in the indirect moxibustion plaster group. The cured and markedly effective rate of the combination group was significantly higher than that of the Chinese medicine group （χ²=4.593， P<0.05） and the indirect moxibustion plaster group （χ²=5.711， P<0.05）. The total effective rate was 96.7 % （29/30） in the combination group， 93.3% （28/30） in the Chinese medicine group， and 86.7% （26/30） in the indirect moxibustion plaster group. The total effective rate of the combination group was higher than that of the Chinese medicine group and the indirect moxibustion plaster group， but the differences were not statistically significant. ConclusionChinese medicine combined with indirect moxibustion plaster can effectively improve the clinical symptoms，promote pulmonary inflammation，blood routine indexes， and inflammatory indexes， and improve the quality of life of COVID-19 patients during the recovery period，which is more advantageous than Chinese medicine alone or indirect moxibustion plaster.

Objective    To investigate the role of kinase insert domain containing receptor (KDR) positive cells in the formation of cardiospheric structure, myocardium and vessels. Methods    Twenty-four Wistar rats weighting 250 g were selected. Cardiosphere-derived cells were isolated by enzymatic digestion of rat hearts, and their immunological phenotypes were analyzed by using fluorescence-activated cell sorting (FACS). The cardiomyogenic and vasculogenic potential was diagnosed by immunohistochemistry. Results    KDR positive cells grew exponentially and formed cell clusters. It also could generate myocardial precursor cells (cardiac troponin T positive). And these cells can develop spontaneous contraction activity in vitro. Meanwhile, KDR positive cells formed many vessel-like structures through a budding process. Conclusion    KDR positive cells form cardiospheric structure in vitro culture, and exhibit differentiation potential towards the cardiac and vascular cells. Therefore, KDR positive cells may have a broad prospect of clinical application as cell donors.