Objective:To discuss the effect of knock down of gene of kinesin family member 3B(KIF3B),an important component of primary cilia(PC)of in mouse embryonic palatal mesenchymal on the autophagy level of cells(mEPMCs)cells,and to clarify its mechanism.Methods:The mEPMCs from gestational day 14.5 C57BL/6J mice cultured in vitro were collected and divided into control group(administered normal saline),empty lentivirus transfected cell group(sh-NC group)(administered lentivirus transfection),KIF3B knockdown group(sh-KIF3B group)(administered KIF3B gene knockdown),and KIF3B knockdown plus Smoothened receptor agonist(SAG)group(sh-KIF3B+SAG group)(administered KIF3B gene knockdown followed by SAG addition),based on whether the KIF3B gene was knocked down and whether the SAG was used to activate the sonic hedgehog(Shh)signaling pathway and its downstream coreceptor Smo,with 5 rats in each group.Transmission electron microscope was used to observe the morphology and the number of autophagosomes/autolysosomes in the mEPMCs in various groups;Western blotting method was used to detect the expression levels of autophagy-related proteins Beclin-1 and p62,and the Shh signaling pathway proteins Shh and Smo in the mEPMCs in various groups.Results:The transmission electron microscope observation results showed that compared with control group,the number of autophagosomes/autolysosomes in sh-KIF3B group was significantly increased(P<0.05);compared with sh-KIF3B group,the number of autophagosomes/autolysosomes in the mEPMCs in sh-KIF3B+SAG group was significantly decreased(P<0.05).The Western blotting results showed that compared with control group,the Beclin-1 protein expression level in the mEPMCs in sh-KIF3B group was significantly increased(P<0.05),and the KIF3B,p62,Shh,and Smo protein expression levels were significantly decreased(P<0.01);compared with sh-KIF3B group,the Shh,Smo,and p62 protein expression levels in the mEPMCs in sh-KIF3B+SAG group were significantly increased(P<0.01),and the Beclin-1 protein expression level was significantly decreased(P<0.01).Conclusion:Knockdown of KIF3B gene can promote autophagy of the mEPMCs,and the mechanism may be related to its inhibition of the Shh signaling pathway.

BACKGROUND:Studies demonstrated that miR-135a-5p was highly expressed in mouse embryonic palatal mesenchymal cells with cleft palate induced by dexamethasone.The primary cilium and its mediated Shh signaling pathway were involved in the autophagy of mouse embryonic palatal mesenchymal cells.It is speculated that miR-135a-5p may regulate autophagy in mouse embryonic palatal mesenchymal cells through primary cilia and its mediated Shh signaling pathway. OBJECTIVE:To investigate the regulatory effect of miR-135a-5p on autophagy of mouse embryonic palatal mesenchymal cells. METHODS:In vitro,palatal mesenchymal cells from C57BL/6J mouse embryos were extracted and cultured.Cell transfections were set up as follows:(1)the cells were divided into control group,miR-135a-5p negative control group and miR-135a-5p mimic group;(2)NC+miR-NC group,KIF3B overexpression group,and miR-135a-5p+KIF3B group:qRT-PCR was performed to verify transfection efficiency of miR-135a-5p and KIF3B.A transmission electron microscope was used to observe the number of autophagosome/autophagolysosome in the cells of each group.The degree of fluorescence expression of autophagy marker LC3B was determined by the immunofluorescence technique.The protein expression of KIF3B,LC3 and P62 was determined by western blot assay.(3)The cells were divided into miR-135a-5p negative control group,and SAG treated group,and SAG+miR-135a-5p group.qRT-PCR was used to detect the mRNA expression levels of Gli3,a key transcription factor downstream of Shh signaling.The protein expressions of autophagy-related proteins LC3 and P62 were detected by western blot assay. RESULTS AND CONCLUSION:(1)After overexpression of miR-135a-5p,the number of autophagosome/autophagolysosome was significantly increased(P<0.01).The fluorescence density of LC3B increased significantly(P<0.01);the protein expression of KIF3B and P62 decreased(P<0.01),and the protein expression of LC3 increased.(2)After overexpression of KIF3B,the number of autophagosome/autophagolysosome was significantly decreased(P<0.01);the fluorescence density of LC3B was decreased(P<0.01);the protein expression of P62 was increased(P<0.01),and the protein expression of LC3 was decreased(P<0.01).Targeted expression of KIF3B was inhibited by miR-135a-5p(P<0.01);the number of autophagosome/autophagolysosome,the fluorescence intensity of LC3B as well as the protein expression of LC3 were reversed(P<0.01)and the protein expression of P62 was decreased(P<0.01).(3)SAG significantly increased the mRNA expression of Gli3(P<0.01),increased the protein expression of P62(P<0.01),and decreased the protein expression of LC3(P<0.01).When miR-135a-5p was added,Gli3 mRNA expression was significantly decreased(P<0.01);P62 protein expression was decreased(P<0.01),and LC3 protein expression was reversed(P<0.01).(4)These results indicate that miR-135a-5p targets the inhibition of KIF3B and promotes autophagy in mouse embryonic mesenchymal cells possibly by negatively regulating the Shh signaling pathway.

Objective To explore the predictive value of cystatin C(Cys C),high mobility group box 1(HMGB1),and growth differentiation factor-15(GDF-15)levels for early infection after flap reconstruction for diabetic foot ulcer(DFU).Methods From July 2021 to March 2024,155 DFU patients treated in our hospital were included in DFU group.DFU patients were assigned into non-infection group(104 cases)and infection group(51 cases)according to whether there was infection at the operation site within one week after flap reconstruction.Control group included 85 patients with diabetes but without foot ulcer.Latex immunoturbidimetry was applied to detect serum Cys C level;enzyme linked immunosorbent assay to detect serum levels of HMGB1 and GDF-15;multivariate logistic regression to analyze the factors affecting early infection after flap reconstructionfor DFU,and receiver operating characteristic(ROC)curve to analyze the predictive value of serum Cys C,HMGB1,and GDF-15 levels for early infection after flap reconstruction for DFU.Results The expression levels of serum Cys C,HMGB1,and GDF-15 were obviously higher in the DFU group(P＜0.05)when compared with those in the control group.The expression levels of FPG,CRP,Cys C,HMGB1,and GDF-15 were obviously higher in the infection group(P＜0.05)when compared with those in the non-infection group.Cys C,HMGB1,GDF-15,FPG,and CRP were all inde-pendent risk factors for early infection after flap reconstruction for DFU(P＜0.05).The AUC predicted by serum Cys C,HMGB1,and GDF-15 alone for early infection after flap reconstruction for DFU was 0.810,0.850,and 0.828,respectively.The AUC predicted by the combination of these three markers was 0.930,which was better than that predicted by the three markers alone(ZCysC-three combination=3.381,ZHMGB1-three combination=2.588,ZGDF-15-three combination=2.857,all P＜0.05).Conclusions Cys C,HMGB1,and GDF-15 are upregulated in the serum of DFU patients,and all the three are factors affecting early infection after flap reconstruction for DFU.The combination of the three has high predictive value for early infection after DFU flap repair.

Autoimmune glial fibrillary acidic protein astropathy(GFAP-A)is an autoimmune inflammatory disease of the central nervous system,with a prevalence rate of 0.6 per 100,000.Its clinical manifestations include subacute meningitis,encephalitis,myelitis,or combinations thereof.Approximately 40%of patients exhibit symptoms and signs of prodromal infection such as fever and headache,while 30%may develop tumors.Currently,diagnosis primarily relies on the presence of GFAP-IgG in cerebrospinal fluid.Differential diagnosis must exclude conditions such as central nervous system vasculitis,inflammatory demyelinating diseases,lymphoma,glioma,and brain metastases,posing significant challenges in clinical practice.This paper summarizes the clinical manifestations,diagnosis,and treatment of GFAP-A to enhance understanding of the disease and prevent misdiagnosis.

Objective To explore the predictive value of cystatin C(Cys C),high mobility group box 1(HMGB1),and growth differentiation factor-15(GDF-15)levels for early infection after flap reconstruction for diabetic foot ulcer(DFU).Methods From July 2021 to March 2024,155 DFU patients treated in our hospital were included in DFU group.DFU patients were assigned into non-infection group(104 cases)and infection group(51 cases)according to whether there was infection at the operation site within one week after flap reconstruction.Control group included 85 patients with diabetes but without foot ulcer.Latex immunoturbidimetry was applied to detect serum Cys C level;enzyme linked immunosorbent assay to detect serum levels of HMGB1 and GDF-15;multivariate logistic regression to analyze the factors affecting early infection after flap reconstructionfor DFU,and receiver operating characteristic(ROC)curve to analyze the predictive value of serum Cys C,HMGB1,and GDF-15 levels for early infection after flap reconstruction for DFU.Results The expression levels of serum Cys C,HMGB1,and GDF-15 were obviously higher in the DFU group(P＜0.05)when compared with those in the control group.The expression levels of FPG,CRP,Cys C,HMGB1,and GDF-15 were obviously higher in the infection group(P＜0.05)when compared with those in the non-infection group.Cys C,HMGB1,GDF-15,FPG,and CRP were all inde-pendent risk factors for early infection after flap reconstruction for DFU(P＜0.05).The AUC predicted by serum Cys C,HMGB1,and GDF-15 alone for early infection after flap reconstruction for DFU was 0.810,0.850,and 0.828,respectively.The AUC predicted by the combination of these three markers was 0.930,which was better than that predicted by the three markers alone(ZCysC-three combination=3.381,ZHMGB1-three combination=2.588,ZGDF-15-three combination=2.857,all P＜0.05).Conclusions Cys C,HMGB1,and GDF-15 are upregulated in the serum of DFU patients,and all the three are factors affecting early infection after flap reconstruction for DFU.The combination of the three has high predictive value for early infection after DFU flap repair.

Autoimmune glial fibrillary acidic protein astropathy(GFAP-A)is an autoimmune inflammatory disease of the central nervous system,with a prevalence rate of 0.6 per 100,000.Its clinical manifestations include subacute meningitis,encephalitis,myelitis,or combinations thereof.Approximately 40%of patients exhibit symptoms and signs of prodromal infection such as fever and headache,while 30%may develop tumors.Currently,diagnosis primarily relies on the presence of GFAP-IgG in cerebrospinal fluid.Differential diagnosis must exclude conditions such as central nervous system vasculitis,inflammatory demyelinating diseases,lymphoma,glioma,and brain metastases,posing significant challenges in clinical practice.This paper summarizes the clinical manifestations,diagnosis,and treatment of GFAP-A to enhance understanding of the disease and prevent misdiagnosis.

Cerebral microbleeds (CMBs) are an imaging biomarker of cerebral small vessel disease (CSVD). Researches have shown that CMBs are a risk factor for hemorrhagic transformation and poor outcomes after reperfusion therapy in patients with acute ischemic stroke. This article reviews the relationship between CMBs and the outcomes of reperfusion therapy in patients with acute ischemic stroke.

The bacterial ATP-competitive GyrB/ParE subunits of type II topoisomerase are important anti-bacterial targets to treat super drug-resistant bacterial infections. Herein we discovered novel pyrrolamide-type GyrB/ParE inhibitors based on the structural modifications of the candidate AZD5099 that was withdrawn from the clinical trials due to safety liabilities such as mitochondrial toxicity. The hydroxyisopropyl pyridazine compound 28 had a significant inhibitory effect on Gyrase (GyrB, IC₅₀ = 49 nmol/L) and a modest inhibitory effect on Topo IV (ParE, IC₅₀ = 1.513 μmol/L) of Staphylococcus aureus. It also had significant antibacterial activities on susceptible and resistant Gram-positive bacteria with a minimum inhibitory concentration (MIC) of less than 0.03 μg/mL, which showed a time-dependent bactericidal effect and low frequencies of spontaneous resistance against S. aureus. Compound 28 had better protective effects than the positive control drugs such as DS-2969 ( 5) and AZD5099 ( 6) in mouse models of sepsis induced by methicillin-resistant Staphylococcus aureus (MRSA) infection. It also showed better bactericidal activities than clinically used vancomycin in the mouse thigh MRSA infection models. Moreover, compound 28 has much lower mitochondrial toxicity than AZD5099 ( 6) as well as excellent therapeutic indexes and pharmacokinetic properties. At present, compound 28 has been evaluated as a pre-clinical drug candidate for the treatment of drug-resistant Gram-positive bacterial infection. On the other hand, compound 28 also has good inhibitory activities against stubborn Gram-negative bacteria such as Escherichia coli (MIC = 1 μg/mL), which is comparable with the most potent pyrrolamide-type GyrB/ParE inhibitors reported recently. In addition, the structure-activity relationships of the compounds were also studied.

Objective:To summarize the age, gender difference, cause, location, time, treatment and prognosis of femoral intertrochanteric fracture complicated with pseudoaneurysm, and analyze the influence of different treatment methods on the occurrence of femoral intertrochanteric fracture complicated with pseudoaneurysm.Methods:Using the method of systematic literature review, a total of 76 literatures were retrieved from multiple databases at home and abroad, and 45 literatures were screened out through the second screening of title, abstract and full text. The patients with intertrochanteric fracture complicated with pseudoaneurysm were selected as the research objects, and the duplicate cases were deleted; the age, gender, onset age, etiology, location, treatment and prognosis of the disease were retrospectively analyzed.Results:A total of 54 cases of femoral intertrochanteric fracture related pseudoaneurysms were selected from 45 literatures, including 25 males and 27 females, and 2 cases had no gender; aged from 43 to 94 years old, and 4 cases were younger than 60 years old, 6 between 61 and 70, 17 between 71 and 80, and 25 between 81 and 90. Forty-five cases occurred in deep femoral artery, 3 cases in superficial femoral artery, 1 case in superior gluteal artery, 1 case of medial circumflex artery, 1 case of lateral circumflex femoral artery, 1 case of femoral artery, and 2 cases of unknown location. The occurrence time of pseudoaneurysm: 17 cases within 1 week after fracture, 8 cases from 1 week to 3 weeks, 19 cases from 3 weeks to 3 months, 2 cases more than 3 months, and 8 cases unknown. The causes of pseudoaneurysm: among the 54 patients, 41 cases had definite causes, including 21 cases of vascular injury caused by fracture block; 20 cases of iatrogenic injury, of which the incidence of direct injury of blood vessels by screw or drill was the highest, accounting for 55% (11/20). Twenty-two cases were treated with vascular embolization, 10 cases with surgical sutures, 8 cases with surgical ligation, 4 cases with resection of pseudoaneurysm, 5 cases with covered stent, and 4 cases with thrombin injection. There were 42 cases who had follow-up data, and no recurrence and other complications were found during the follow-up period.Conclusion:The peak age of femoral intertrochanteric fracture complicated with pseudoaneurysm was 71-90 years old. The incidence of femoral intertrochanteric fracture complicated with pseudoaneurysm was more often within 1 week after fracture and 3 weeks to 3 months after fracture, due to fracture fragments and iatrogenic vascular injury caused by the location of the profundus femoris artery. Treatment options include vascular embolization, surgical ligation, suture or resection of pseudoaneurysm, covered stent and thrombin injection, and the prognosis is generally good.

Inflammatory bowel disease (IBD) is a chronic and recurrent inflammatory disease of gastrointestinal tract, including Crohn's disease and ulcerative colitis. The exact etiology and pathogenesis of IBD are still not clear. Neutrophil gelatinase-associated lipocalin (NGAL) plays an important role in regulating intestinal immunity, maintaining intestinal epithelial cell barrier, and coordinating the composition of intestinal flora. In addition, NGAL is useful for helping the clinical testing of IBD. This article reviewed the research progress of NGAL in IBD.