[Objective]To investigate the effect and mechanism of Shuxuetong and its main component hirudin on the apoptosis of cerebellar granule neurons(CGNs)in Sprague-Dawley(SD)rats.[Methods]CGNs incubated in vitro for 7 days were divided into survival control group or 25 K group(cultured in medium containing 25 mmol/L KCL)and apopto-sis group or 5 K group(cultured in medium containing 5 mmol/L KCL).CGNs were separately treated with proportionally diluted and different concentrations of Shuxuetong(1/50,1/40,1/30,1/20 and 1/10)and the corresponding different con-centrations of hirudin(2,2.5,3.34,5 and 10 U/mL).Hoechst staining was performed to analyze the apoptosis.Western blot was used to detect the expression levels of Cleaved Caspase-3,Bim and VEGF.[Results]Hoechst staining showed that 5 K group had a higher apoptosis rate than 25 K group.In 25 K group,there was no significant change in the apoptosis rate between neurons treated with different concentrations of Shuxuetong and hirudin,but significant changes was found in 5 K group and the higher the concentration,the lower the apoptosis rate.Western blot results revealed that,compared with control neurons in 5 K group,Shuxuetong injection and hirudin treatments resulted in a decrease of Cleaved Caspase-3 and Bim expression,but an increase of VEGF protein.[Conclusions]Shuxuetong and its main component hirudin inhibits the apoptosis of CGNs through suppressing proapoptotic BH3-only protein Bim.

Ankylosing spondylitis (AS) combined with lower cervical fracture is often categorized into unstable fracture, with a high incidence of neurological injury and a high rate of disability and morbidity. As factors such as shoulder occlusion may affect the accuracy of X-ray imaging diagnosis, it is often easily misdiagnosed at the primary diagnosis. Non-operative treatment has complications such as bone nonunion and the possibility of secondary neurological damage, while the timing, access and choice of surgical treatment are still controversial. Currently, there are no clinical practice guidelines for the treatment of AS combined with lower cervical fracture with or without dislocation. To this end, the Spinal Trauma Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults ( version 2024) in accordance with the principles of evidence-based medicine, scientificity and practicality, in which 11 recommendations were put forward in terms of the diagnosis, imaging evaluation, typing and treatment, etc, to provide guidance for the diagnosis and treatment of AS combined with lower cervical fracture.

Diabetes mellitus(DM)is a disease syndrome characterized by chronic hyperglycaemia.A long-term high-glucose environment leads to reactive oxygen species(ROS)production and nuclear DNA damage.Human umbilical cord mesenchymal stem cell(HUcMSC)infusion induces significant antidiabetic effects in type 2 diabetes mellitus(T2DM)rats.Insulin-like growth factor 1(IGF1)receptor(IGF1R)is important in promoting glucose metabolism in diabetes;however,the mechanism by which HUcMSC can treat diabetes through IGF1R and DNA damage repair remains unclear.In this study,a DM rat model was induced with high-fat diet feeding and streptozotocin(STZ)administration and rats were infused four times with HUcMSC.Blood glucose,interleukin-6(IL-6),IL-10,glomerular basement membrane,and renal function were examined.Proteins that interacted with IGF1R were determined through coimmunoprecipitation assays.The expression of IGF1R,phosphorylated checkpoint kinase 2(p-CHK2),and phosphorylated protein 53(p-p53)was examined using immunohistochemistry(IHC)and western blot analysis.Enzyme-linked immunosorbent assay(ELISA)was used to determine the serum levels of 8-hydroxydeoxyguanosine(8-OHdG).Flow cytometry experiments were used to detect the surface markers of HUcMSC.The identification of the morphology and phenotype of HUcMSC was performed by way of oil red"O"staining and Alizarin red staining.DM rats exhibited abnormal blood glucose and IL-6/10 levels and renal function changes in the glomerular basement membrane,increased the expression of IGF1 and IGF1R.IGF1R interacted with CHK2,and the expression of p-CHK2 was significantly decreased in IGF1R-knockdown cells.When cisplatin was used to induce DNA damage,the expression of p-CHK2 was higher than that in the IGF1R-knockdown group without cisplatin treatment.HUcMSC infusion ameliorated abnormalities and preserved kidney structure and function in DM rats.The expression of IGF1,IGF1R,p-CHK2,and p-p53,and the level of 8-OHdG in the DM group increased significantly compared with those in the control group,and decreased after HUcMSC treatment.Our results suggested that IGF1R could interact with CHK2 and mediate DNA damage.HUcMSC infusion protected against kidney injury in DM rats.The underlying mechanisms may include HUcMSC-mediated enhancement of diabetes treatment via the IGF1R-CHK2-p53 signalling pathway.

Osteochondral lesion of talus (OLT) is a foot and ankle disease characterized by ankle pain, which may impact the joint function and life quality. If managed improperly, it may lead to a further ankle arthritis, severely compromising the prognosis. The therapeutic effect of conservative treatment for OLT is still uncertain. Surgery is still the main treatment modality for OLT with various techniques. However, the optimized surgical technique is still inconclusive, furthermore, regeneration and repair of cartilage after debridement is also a great challenge for the treatment of OLT. Platelet-rich plasma (PRP) with good repair effect on cartilage injury is gradually applied in the treatment of OLT. However, there still lacks the unified understanding of the technique and specification of PRP for the treatment of OLT. Therefore, National Orthopedics Center of Shanghai Sixth People′s Hospital allied Foot Ankle Basic Research & Orthopedics Group, Chinese Association of Orthopedic Surgeons; Foot and Ankle Committee of Chinese Association of Sports Medicine Physicians; and Foot and Ankle Group of Orthopedic Specialized Branch of Shanghai Medical Association to organize related experts to formulate the Expert consensus on platelet- rich plasma treatment for osteochondral lesion of talus ( version2023). Fifteen recommendations were put forward upon PRP preparation, indications, contraindications and treatment methods of PRP for OLT, so as to standardize the PRP treatment for OLT.

Lower extremity deep vein thrombosis (DVT) is one of the main complications in patients with traumatic fractures, and for severe patients, the DVT can even affect arterial blood supply, resulting in insufficient limb blood supply. If the thrombus breaks off, pulmonary embolism may occur, with a high mortality. The treatment and rehabilitation strategies of thrombosis in patients with lower extremity fractures have its particularity. DVT in traumatic fractures patients has attracted extensive attention and been largely studied, and the measures for prevention and treatment of DVT are constantly developing. In recent years, a series of thrombosis prevention and treatment guidelines have been updated at home and abroad, but there are still many doubts about the prevention and treatment of DVT in patients with different traumatic fractures. Accordingly, on the basis of summarizing the latest evidence-based medical evidence at home and abroad and the clinical experience of the majority of experts, the authors summarize the clinical treatment and prevention protocols for DVT in patients with traumatic fractures, and make this consensus on the examination and assessment, treatment, prevention and preventive measures for DVT in patients with different fractures so as to provide a practicable approach suitable for China ′s national conditions and improve the prognosis and the life quality of patients.

Objective To investigate the effect of microRNA-96-5p(miR-96-5p)on expression of mam-malian target of rapamycin(mTOR)in gastric cancer cells. Methods Lentiviral vector,LV-hsa-mir-96-5p was transfected into BGC-823 gastric cancer cells. The expression of miR-96-5p and mTOR mRNA after transfection was detected by qRT-PCR,and the expression of mTOR protein was detected by Western blot. Results At 72 h after transfection,the expression level of miR-96-5p in the experimental group was 20.4 times of that in the blank control group(P<0.05),while the expression level of mTOR gene was downregulated to 0.47 times of that in the blank control group (P < 0.05),and mTOR protein relative expression was significantly reduced compared with the control group (P < 0.05). Conclusion Overexpression of miR-96-5p can downregulate the expression of mRNA and protein of mTOR gene in gastric cancer cells,thus,mTOR may be one of the downstream target genes of miR-96-5p.

AIM:To study whether inhibition of forkhead box protein M1(FoxM1) sensitizes leukemia K562 cells to homoharringtonine ( HHT ) .METHODS: K562 cells were incubated with HHT at different concentrations ( 0μmol/L, 0.015 μmol/L, 0.030μmol/L and 0.045μmol/L) for different time (0 h, 24 h, 48 h and 72 h).The mRNA and protein levels of FoxM1 were detected by real-time PCR and Western blot.FoxM1 siRNA was transfected into K562 cells with 0.015μmol/L HHT after 6 h.After 72 h incubation, the cell proliferation was detected by cell counting and soft agar assay, and the proportion of apoptotic K562 cells was determined by flow cytometry.The expression of c-Myc and Sp1 were detected by real-time PCR and Western blot.RESULTS:FoxM1 expression was reduced time-dependently and dose-dependently, suggesting that HHT mediated the downregulation of FoxM1 in K562 cells.In K562 cells, treatment with FoxM1 siRNA and HHT inhibited the cell proliferation and promoted the apoptosis significantly.Therefore, inhibition of FoxM1 sensitized leukemia K562 cells to HHT.The expression of c-Myc and Sp1 was positively regulated by FoxM1. CONCLUSION:HHT inhibits Forkhead box protein M1 expression in K562 cells.Inhibition of FoxM1 sensitizes K562 cells to HHT.

Objective To investigate the mechanism of histone deacetylase inhibitors in proliferation ofglioma cells.Methods (1) Glioma cell line U251 was cultured in vitro and treated with Trichostatin A (TSA) at 0.1,0.2,0.5,1.0 or 2.0 μmol/L for 48 h to determine the IC50 for TSA,and then,cells were treated with TSA at the IC50 concentration (0.5 μ mol/L) for 8,16,24,48 h;1 μmol/L M344,0.5 μmol/L LBH589,6 mmol/L NaBu and 6 mmol/L VPA were used to treat the cells for 48 h and same volume of solvent was given to cells for 48 h as control group;MTT assay was performed to determine cell viability.(2) Western blotting was performed to test the Jun N-terminal kinase (JNK)expression and phosphorylated JNK (p-JNK) level in the U251 cells after being treated with 0.5 μmol/L TSA for 8,16 and 24 h.(3) Western blotting and MTT assay were employed to detect the c-Jun expression and phosphorylated c-Jun (p-c-Jun) level,and cell viability in the U251 cells of control group,10 μmol/L SP600125 treatment group (JNK inhibitor) and 1 μmol/L CEP11004 treatment group (MLK3,a direct upstream kinase of JNK).(4) Western blotting and MTT assay were employed to detect the c-Jun and Flag expressions,and cell viability in the U251 cells of pcDNA3.1 transfected group,pcDNA3.1+TSA transfected group,pMKK7-JNK1 transfected group and pMKK7-JNK1+TSA transfected group.Results (1) The viability of cells of 0.1,0.2,0.5,1.0 or 2.0 μmol/L TSA treated group was significantly lower than that in the control group,and the higher the TSA concentration,the lower the viability of cells;the viability of cells of 0.5 μmol/L TSA treated for 16,24,36 and 48 h groups was significantly lower than that in the control group,and the longer the TSA treatment,the lower the viability of cells.(2) The viability of 1 μmol/L M344,0.5 μmol/L LBH589,6 mmol/L NaBu and 6 mmol/L VPA treatment groups was significantly lower than that in the control group (P＜0.05).(3) As compared with those in the control group,the p-JNK expression was significantly decreased in the cells after 0.5 μmol/L TSA treatment for 16 and 24 h,and the p-c-Jun protein expression and the cell viability were significantly decreased in the cells of 10 μmol/L SP600125 and 1 μmol/L CEP11004 treatment groups (P＜0.05).(4) As compared with that in the pcDNA3.1 transfected group,cell viability in the U251 cells ofpcDNA3.1+TSA transfected group,pMKK7-JNK1 transfected group and pMKK7-JNK1+TSA transfected group was significantly increased(P＜0.05).Conclusion Histone deacetylase inhibitors reduce the proliferation of glioma cell U251 through inhibiting JNK activity.

Objective To evaluate the changes of the level of cerebrospinal fluid T-tau protein andβ-amyloid 42 in mild cognitive impairment (MCI) patients. Methods Computer retrieval was preformed for cerebrospinal fluid (CSF) to-tal tau protein (T-tau) and β-amyloid 42 (Aβ 42) levels among MCI and healthy people in Pubmed, Cochrane Library, Ovid, CNKI, VIP Data and Wangfang Data. Meta-analyses were conducted on the T-tau and Aβ42 levels using Review Manager 5.2 software. Results Seventeen studies met inclusion criteria. Meta-analysis results showed that, CSF T-tau protein levels were significantly higher (MD:113.10;95%CI:87.93-138.26) and Aβ42 levels were significantly lower in the patients with MCI, (MD=-146.50;95%CI=-178.70- -114.29) compared with the control group (P<0.01). Conclu-sions T-tau significantly increases while Aβ42 significantly decreases in the CSF in patients with MCI.

Objective To analysis the malignant performance characteristics of tumor stem cell-like side popula-tion cells in patients with cervical cancer. Methods The cervical cancer cells were obtained from surgical resection tumor tissue. The tumor stem cell-like side population cells were isolated by flow cytometry. The cell growth curve was drawn by MTT assay. The invasion ability of tumor cells was compared by transwell assay. The clonogenic capacity was detected by clone formation in soft agar. The expression level of ABCG2 protein, a drug-resistant gene, was detected by immunofluores-cence method. Finally, these cells were transplated into the subcutaneous of de thymus mice. The rate of tumor formation was compared between groups. Results The results from flow cytometry assay showed the percentage of cervical cancer stem cell-like side population cells was 1.39%. Compared with the non-side population cells, the side population cells grow quickly, showed the enhanced invasion ability and colony forming ability. There was more high expression level in ABCG 2 protein of side population cells. The tumor form rate was 100%(10/10) in the side population cells and the non-side popula-tion cells was 20%(2/10). Conclusion The cervical cancer stem cell-like side population cells have more malignant perfor-mance characteristics than that of non-side population cells, which maybe a core target for cancer gene therapy in the future.