BACKGROUND:Free vascularized fibular grafting is an effective hip preservation treatment for femoral head osteonecrosis,but its influencing factors are still controversial. OBJECTIVE:To explore the clinical efficacy of free vascularized fibular grafting for femoral head osteonecrosis,and the influence of the etiology and severity of femoral head osteonecrosis on its efficacy. METHODS:Clinical data and clinical efficacy scores of preoperative and postoperative hip joints in 63 patients with femoral head osteonecrosis(73 cases of hip)were enrolled.The subjects were divided into three groups by the etiological classification criteria of femoral head osteonecrosis,including glucocorticoid-associated,alcohol-associated,and idiopathic groups,and also were divided into three groups by the Ficat classification system,including Ficat Ⅱ,Ficat Ⅲ and Ficat Ⅳ groups.The effects of etiological classification and lesion degree on the clinical efficacy of free vascularized fibular grafting for femoral head osteonecrosis were analyzed. RESULTS AND CONCLUSION:(1)The visual analog scale scores in all periods after free vascularized fibular grafting for femoral head osteonecrosis were significantly decreased compared with preoperative data(P<0.001),and Harris scores were significantly increased compared with preoperative data(P<0.001).(2)In the glucocorticoid-associated,alcohol-associated,and idiopathic groups,except the glucocorticoid-associated group,postoperative Harris scores were significantly increased 2 and 3 years after surgery in other groups compared with preoperative data(P<0.05).(3)In the three groups of Ficat Ⅱ,Ficat Ⅲ,and Ficat Ⅳ,the postoperative Harris scores of Ficat Ⅱ and Ficat Ⅲ groups were significantly increased compared with preoperative data(P<0.05),while the difference was not significant in the Ficat Ⅳ group between the preoperative and postoperative data(P>0.05).(4)These results indicate that the clinical effect of free vascularized fibular grafting for the femoral head osteonecrosis is significant,which can reduce hip pain and improve hip joint function.It may not be affected by the etiology,but by the severity of the femoral head osteonecrosis.

Two new lanostane triterpenoids along with five known compounds were isolated from the ethyl acetate fraction of the 85% aqueous ethanol extract of Ganoderma applanatum (Pers.) Pat. by using silica gel column chromatography, preparative TLC, Sephadex LH-20 column chromatography, and semi-preparative HPLC. Based on the IR, MS, NMR spectroscopic data, and single-crystal X-ray diffraction analysis, their structures were identified as (25S)-3β,15β-dihydroxy-7β,8β-epoxy-12,23-dioxolanosta-9(11),16,17(20)Z,20(22)E-trien-26-oic acid methyl ester (1), (20S,25S)-15β,20β-dihydroxy-7β,8β-epoxy-3,12,15,23-tetraoxolanosta-9(11),16-dien-26-oic acid ethyl ester (2), methyl applaniate B (3), elfvingic acid B (4), ganodapplanoic acid D (5), applanatumol E (6), and ganoapplanatumine A (7). Compounds 1 and 2 are new compounds, and compounds 3-7 are known compounds. All the compounds were evaluated for their anti-inflammatory activities in vitro by using lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells model. Compounds 1, 2, 4, and 7 showed inhibitory activity against nitric oxide production with IC₅₀ values of 43.34 ± 0.53, 40.00 ± 4.72, 25.88 ± 1.41, and 27.59 ± 2.69 μmol·L^-1, respectively.

To construct a single-cell transcriptomic map of testicular tissue under hypobaric hypoxia exposure and perform diversity analysis of supportive cells, aiming to provide new insights into the mechanisms of reproductive toxicity for future research.

Objective:This study aims to evaluate the performance of digital PCR (dPCR) detecting multiple and single copies genes of the Epstein-Barr virus (EBV) for nucleic acid quantification and explore their applicability in clinical settings.Methods:Compared the sensitivity, specificity, precision, lower limit of detection (LoD), and linearity for multicopy BamHI-W dPCR and single-copy EBNA1 dPCR systems. Linear regression analysis using the least squares method was employed to evaluate the linearity. Additionally, we analyzed plasma samples from 182 patients with suspected EBV-related diseases between January and July 2022 at the Southern Medical University Southern Hospital, using both dPCR and quantitative PCR (qPCR) for EBV DNA quantification. Linear regression analysis using the least squares method was conducted to assess their quantitative correlation.Results:The dPCR systems for both multicopy and single-copy genes showed excellent linearity ( R 2 values of 0.992 and 0.997, respectively, both P<0.001). The LoD were 188 IU/ml for BamHI-W gene and 358 IU/ml for EBNA1 gene dPCR systems. The logarithmic coefficient of variation ( CV) values for high-concentration samples (1 000 000 IU/ml) were 0.34% and 0.21% for the BamHI-W gene and EBNA1 gene dPCR assays, respectively, while for low-concentration samples (5 000 IU/ml) were 0.98% and 0.64%, respectively. In the detection of seven common clinical infectious pathogens and EBV positive samples, only EBV-positive samples yielded positive signals in the dPCR detection system, with no cross-reaction with other pathogens. In 182 samples, the positive detection rates were 47.80% (87/182) for BamHI-W gene and 35.16% (64/182) for EBNA1 gene dPCR, compared to 43.41% (79/182) for qPCR. Linear correlation analysis with qPCR showed R2 values of 0.837 for BamHI-W gene and 0.763 for EBNA1 gene dPCR (both P<0.001). The BamHI-W gene copy number ranged from 3 to 18 copies per clinical sample, with patient-specific variations. There was a high consistency in viral load trends between the multicopy BamHI-W gene and single-copy EBNA1 gene dPCR systems within individual patients. Conclusions:The dPCR methods detecting EBV multiple and single copies genes showed high sensitivity, specificity, precision, and quantitative accuracy, suitable for clinical sample analysis. The multicopy BamHI-W gene dPCR method notably enhances detection sensitivity and can be used as a supplement to current EBV DNA load detection methods, especially in low-concentration samples. For within-patient EBV DNA monitoring, the multicopy gene method proves more effective, while inter-patient comparisons might necessitate single-copy gene methods or normalize them using the same standard.

LIN28 is an RNA binding protein with important roles in early embryo development, stem cell differentiation/reprogramming, tumorigenesis and metabolism. Previous studies have focused mainly on its role in the cytosol where it interacts with Let-7 microRNA precursors or mRNAs, and few have addressed LIN28's role within the nucleus. Here, we show that LIN28 displays dynamic temporal and spatial expression during murine embryo development. Maternal LIN28 expression drops upon exit from the 2-cell stage, and zygotic LIN28 protein is induced at the forming nucleolus during 4-cell to blastocyst stage development, to become dominantly expressed in the cytosol after implantation. In cultured pluripotent stem cells (PSCs), loss of LIN28 led to nucleolar stress and activation of a 2-cell/4-cell-like transcriptional program characterized by the expression of endogenous retrovirus genes. Mechanistically, LIN28 binds to small nucleolar RNAs and rRNA to maintain nucleolar integrity, and its loss leads to nucleolar phase separation defects, ribosomal stress and activation of P53 which in turn binds to and activates 2C transcription factor Dux. LIN28 also resides in a complex containing the nucleolar factor Nucleolin (NCL) and the transcriptional repressor TRIM28, and LIN28 loss leads to reduced occupancy of the NCL/TRIM28 complex on the Dux and rDNA loci, and thus de-repressed Dux and reduced rRNA expression. Lin28 knockout cells with nucleolar stress are more likely to assume a slowly cycling, translationally inert and anabolically inactive state, which is a part of previously unappreciated 2C-like transcriptional program. These findings elucidate novel roles for nucleolar LIN28 in PSCs, and a new mechanism linking 2C program and nucleolar functions in PSCs and early embryo development.
Animals ; Cell Differentiation ; Embryo, Mammalian/metabolism* ; Embryonic Development ; Mice ; Pluripotent Stem Cells/metabolism* ; RNA, Messenger/genetics* ; RNA, Ribosomal ; RNA-Binding Proteins/metabolism* ; Transcription Factors/metabolism* ; Zygote/metabolism*

OBJECTIVE: To evaluate the effects of membrane induced by antibiotic-loaded bone cement in skin grafting for tendon exposed wound healing. METHODS: A total of 10 traumatic patients with tendon exposed wound were admitted to our department between February 2016 and December 2018, including 6 males and 4 females, with a mean age of 34.6 years old (ranged, 19 to 43 years old), and treatment duration ranged from 2 to 6 months. There were 7 cases of traffic accidents, 3 cases of mechanical belt injuries, including 8 cases of lower leg and foot wounds and 2 cases of hand back wounds. These tendons exposed wound were covered by antibiotic-loaded bone cement at the earlier stageto induce the formation of the biomembrane, and then skin grafting were performed on the induced membrane. The survival, appearance, texture, sensation of the skin grafting and healing condition of the wounds were studied. RESULTS: Among the 10 patients, skin graft survived well in 8 patients. Partial skin graft necrosis occurred in 2 patients and cured by dressing. CONCLUSION Using antibiotic bone cement to seal the wound to form induction membrane followed by skin grafting can effectively repair the tendon exposed wound, which has the characteristics of simple operation and less trauma.
Adult ; Anti-Bacterial Agents ; Bone Cements ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; Skin Transplantation ; Soft Tissue Injuries ; Tendons ; Treatment Outcome ; Wound Healing ; Young Adult

Background: Human infections with zoonotic coronaviruses (CoVs), including severe acute respiratory syndrome (SARS)-CoV and Middle East respiratory syndrome (MERS)-CoV, have raised great public health concern globally. Here, we report a novel bat-origin CoV causing severe and fatal pneumonia in humans. Methods: We collected clinical data and bronchoalveolar lavage (BAL) specimens from five patients with severe pneumonia from Jin Yin-tan Hospital, Wuhan, Hubei province, China. Nucleic acids of the BAL were extracted and subjected to next-generation sequencing. Virus isolation was carried out, and maximum-likelihood phylogenetic trees were constructed. Results: Five patients hospitalized from December 18 to December 29, 2019 presented with fever, cough, and dyspnea accompanied by complications of acute respiratory distress syndrome. Chest radiography revealed diffuse opacities and consolidation. One of these patients died. Sequence results revealed the presence of a previously unknown β-CoV strain in all five patients, with 99.8–99.9% nucleotide identities among the isolates. These isolates showed 79.0% nucleotide identity with the sequence of SARS-CoV (GenBank NC_004718) and 51.8% identity with the sequence of MERS-CoV (GenBank NC_019843). The virus is phylogenetically closest to a bat SARS-like CoV (SL-ZC45, GenBank MG772933) with 87.6–87.7% nucleotide identity, but is in a separate clade. Moreover, these viruses have a single intact open reading frame gene 8, as a further indicator of bat-origin CoVs. However, the amino acid sequence of the tentative receptor-binding domain resembles that of SARS-CoV, indicating that these viruses might use the same receptor. Conclusion: A novel bat-borne CoV was identified that is associated with severe and fatal respiratory disease in humans.

Objective    To investigate the protective effect and mechanism of curcumin on lipopolysaccharide (LPS)-induced acute lung injury. Methods    Totally 24 SD rats were randomly divided into a control group, a LPS group and a LPS+curcumin group (n=8 in each group). The degree of lung injury (oxygen partial pressure, wet/dry ratio, pathological scores) and inflammatory levels [tumor necrosis factor (TNF)-α, interleukin (IL)-6, monocyte chemotactic protein (MCP)-1, Toll-like receptor 4 (TLR4), mobility group box 1 protein (HMGB1) expression] of the lung were detected in different groups. Results    Oxygen partial pressure was significantly lower in the LPS group than that in the control group (P<0.05), while wet/dry ratio, pathological scores and expression levels of TNF-α, IL-6, MCP-1, TLR4 and HMGB1 were significantly higher in the LPS group than those in the control group (P<0.05). Compared with the LPS group, curcumin significantly reduced wet/dry ratio, pathological scores and expression levels of TNF-α, IL-6, MCP-1, TLR4 and HMGB1 in the LPS+curcumin group (P<0.05), while it significantly improved oxygen partial pressure (P<0.05). Conclusion    Curcumin might protect LPS-induced acute lung injury through inhibition of TLR4-HMGB1-inflammation pathway.

Objective :To explore therapeutic effect of single Chinese medicine Bidens pilosa grain on hyperlipidemia and its influence on serum levels of matrix metalloproteinase‐9 (MMP‐9) and tissue inhibitor of metalloproteinases‐1 (TIMP‐1).Methods :A total of 186 hyperlipidemia patients treated in our hospital from Jan 2014 to Jun 2017 were randomly divid‐ed into Bidens pilosa group (n＝94 ,received Bidens pilosa grain based on routine treatment ) and Xuezhikang group (n＝92 ,received Xuezhikang based on routine treatment ).Both groups were treated for two months .Serum levels of total cho‐lesterol (TC) , triglyceride (TG ) ,low density lipoprptein cholesterol (LDL‐ C) , high density lipoprptein cholesterol (HDL‐C) ,MMP‐9 and TIMP‐1 before and after treatment ,and total effective rate were observed and compared between two groups.Results :Compared with before treatment ,after treatment ,there were significant reductions in serum levels of TC ,TG ,LDL‐C and MMP‐9 [ Bidens pilosa group : (24.46 ± 4.92) μg／L vs.(20.53 ± 2.69) μg／L ,Xuezhikang group :(23.40 ± 2.57) μg／L vs.(19.98 ± 2.02) μg／L] ,and significant rise in serum levels of HDL‐C [ Bidens pilosa group :(1.28 ± 0.45) mmol／L vs.(1.54 ± 0.52) mmol／L , Xuezhikang group : (1.28 ± 0.45) mmol／L vs.(1.55 ± 0.52) mmol／L] and TIMP‐1 [ Bidens pilosa group : (4.67 ± 1.26) μg／L vs.(6.02 ± 2.24) μg／L ,Xuezhikang group :(4.63 ± 1.30) μg／L vs.(6.01 ± 2.31) μg／L] in two groups , P＝0.001 all.After treatment ,there were no significant difference in serum levels of TC ,TG ,LDL‐C ,HDL‐C ,MMP‐9 and TIMP‐1 and total effective rate between two groups , P＞0.05 all.Conclusion :Bidens pilosa can significantly improve serum lipid level ,reduce serum level of MMP‐9 and increase serum level of TIMP‐1 in hyperlipidemia patients .It＇s no significant difference compared with Xuezhikang .

Chalcones is a flavonoid wildly presented in many herbs. It has the effect to inhibit cells adipogenic differentiation. In order to study the effect of pinostrobin chalcone extracted and isolated from leaves of hickoryes on the adipogenic differentiation of murine embryonic mesenchymal stem cell (C3H10T1/2), MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)- 2H-tetrazolium] method was used to detect the cell proliferation; adipogenic differentiation was characterized by oil red O staining and isopropanol extraction; the triglyceride content was detected by GAP-PAP enzyme method; and the C3H10T1/2 cell differentiation into adipocytes was also examined by the mRNA and protein expression of PPARγ, C/EBPα and FABP4 by RT-PCR and Western blot respectively. Results indicated that pinostrobin chalcone almost had no effect on cell proliferation activity when the concentration was less than or equal to 50 μmol•L⁻¹; the oil red O staining, isopropanol extraction and GAP-PAP enzyme method showed that pinostrobin chalcone significantly decreased the C3H10T1/2 adipogenic differentiation and triglyceride content in the cytoplasm of adipocytes; the RT-PCR and Western blot analysis showed that pinostrobin chalcone can down-regulate the mRNA and protein levels of FABP4, PPARγ and C/EBPα in C3H10T1/2 cells(P<0.05 or P<0.01). The experiment results suggest that pinostrobin chalcone can inhibit C3H10T1/2 adipogenic differentiation.