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.

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.

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.

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.

Background: The prevalence of inflammatory bowel disease (IBD) is increasing worldwide, and anemia, one of the most prevalent extraintestinal manifestations of IBD, is still often underdiagnosed and undertreated. Aims: To analyze the correlations of clinical factors with anemia in patients with ulcerative colitis (UC). Methods: A retrospective analysis was conducted in 60 cases of UC admitted from September 2019 to May 2020 at the Second Affiliated Hospital of Zhengzhou University. Sixty healthy subjects were served as controls. Data of fasting venous blood sample analysis on admission were collected, and the correlations of indicators for anemia with the clinical factors were analyzed. Results: The indicators for anemia, including red blood cell (RBC) count, hemoglobin (Hb), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) etc. were significantly decreased in UC patients than in healthy controls (all P<0.05). RBC was lower in females than in males (P<0.05); RBC and HCT were lower in patients with pancolitis than in those with proctitis or left hemi-colon colitis (all P<0.05); RBC, Hb and HCT were lower in severe active UC than in mild active UC (all P<0.05). RBC, Hb, HCT and MCH were negatively correlated with two disease activity indicators for IBD, C-reactive protein (CRP) and/or erythrocyte sedimentation rate (ESR) (all P<0.05). Conclusions: Gender, as well as the disease extent, activity and severity are important clinical factors for anemia in patients with UC.

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.

Objective:To investigate the effect of mechano-growth factor（MGF）on osteoclast activity and its mechanism.Methods:The RAW264.7 precursor osteoclast cell line was cultured with 25 ng/ml macrophage-colony stimulating factor（M-CSF）and 30 ng/ml receptor activator of NF-κB ligand（RANKL），and identified by tartrate resistant acid phosphatase（TRAP）staining after 7 days of culture. Western blot anslysis was used to determine the effect of 45 ng/ml MGF on the phosphoinositide-3-kinase/protein kinase B（PI3K/AKT）signaling pathway in separated osteoclasts，including levels of AKT，phosphorylation（p）-AKT，lactation mammalian target of rapamycin（mTOR），p-mTOR and TRAP at 0，4，8 and 12 hours. Real-time fluorescence quantitative PCR was used to expressions of TRAP in osteoclasts at 0，4，8 and 12 hours. The PI3K/Akt phosphorylation inhibitor LY294002（20 μmol/L）combined with MGF（45 ng/ml）was used to act on osteoclasts，and expression levels of Akt，p-Akt，mTOR，p-mTOR and TRAP were detected by Western blot at 0，4，8 and 12 hours.Results:After culturing RAW264.7 cells with M-CSF and RANKL for 7 days，a large number of osteoclasts with positive TRAP staining can be obtained. Western blot analysis showed expression levels of Akt and mTOR did not change significantly over time（ P>0.05），expression levels of p-Akt and p-mTOR increased continuously from（2.18±0.34）pg/ml and（0.83±0.10）pg/ml at 0 hour to（3.86±0.36）pg/ml and（1.56±0.19）pg/ml at 12 hours（ P<0.05），and expression level of TRAP decreased significantly over time，from（5.66±0.47）pg/ml at 0 hour to（3.76±0.38）pg/ml at 12 hours（ P<0.05）. Real-time fluorescence quantitative PCR analysis of expression of TRAP in osteoclasts showed that MGF inhibited the expression of TRAP in osteoclasts，which decreased from 1.02±0.06 at 0 hour to 0.53±0.11 at 12 hours（ P<0.05）. After acting LY294002 combined with MGF on osteoclasts，Western blot analysis showed expression levels of Akt and mTOR did not change significantly over time（ P>0.05），expression levels of p-AKT and p-mTOR decreased significantly from（3.28±0.18）pg/ml and（3.29±0.22）pg/ml at 0 hour to（2.06±0.34）pg/ml and（2.04±0.20）pg/ml at 12 hours（ P<0.05），and expression level of TRAP had no significant difference over time（ P>0.05）. Conclusions:MGF inhibits osteoclast activity by inhibiting the expression of TRAP in osteoclasts through PI3K/Akt signaling pathway. LY294002 inhibits the expression of PI3K/Akt signaling pathway in osteoclasts，further verifying the mechanism of MGF inhibiting osteoclast activity，and this finding puts forward new ideas for clinical prevention and treatment of osteoporosis.

Exosomes are extracellular vesicles with a diameter of 30-150 nm, which exist in multi-vesicular bodies in the form of intraluminal vesicles. Exosomes can be secreted by a variety of immune cells. Studies have shown that exosomes might play an important role in IBD through their components, such as annexin-A1 (ANXA1), the miRNAs and lipids. As a carrier of antigen presentation, it can affect the signaling pathways related to IBD, regulate the immune response and intestinal homeostasis. This article reviewed the relationship between exosomes and IBD.

MicroRNAs (miRNAs) are a group of short non-coding RNAs consisting of about 22 nucleotides that downregulate gene expression at posttranscriptional level through binding to the 3'-untranslated region (3'-UTR) of target mRNAs. With the increase in research about miRNAs, it is found that miRNAs could modulate the differentiation and secretion of immunocytes as well as the function of intestinal mucosal barrier. Besides, miRNAs could regulate the composition of intestinal microbiome, and could be used as crucial biomarkers for the diagnosis of inflammatory bowel disease (IBD). In this paper, the roles of miRNAs in the pathogenesis of IBD were reviewed through intestinal immunity and barrier function.

Objective: To evaluate the treatment of complex acetabular fractures involving the posterior column by anterograde lag screws via the ilioinguinal approach. Methods: A retrospective study was conducted of the 8 patients with complex acetabular fracture involving the posterior column who had been treated at Department of Orthopaedics, The Second Affiliated Hospital to Inner Mongolia Medical University from January 2017 to June 2018. They were 5 males and 3 females, aged from 35 to 62 years (average, 43.5 years). According to the Letournel-Judet classification, 3 cases were T-shaped fractures, 4 anterior column plus posterior hemitransverse fractures and one both column fracture. The interval from injury to operation averaged 8 days (from 7 to 17 days). The anterior acetabulum was fixated by a reconstruction plate and the posterior column by antegrade lag screws, all through the ilioinguinal approach. The quality of fracture reduction, fracture union time, function of the affected hip and complications were recorded. Results: By the Matta imaging criteria, the quality of fracture reduction was rated as excellent in 7 cases and as fine in one. Intraoperative major hemorrhage or injury to sciatic nerve occurred in none of the patients. This cohort obtained an average follow-up of 8 months (from 6 to 18 months). All fractures united well after an average of 10 weeks (from 8 to 12 weeks). The function of affected hip evaluated by the improved Merle d’Aubigne & Postel criteria at the last follow-up was excellent in 7 cases and fine in one. Follow-ups revealed no incidence of deep vein thrombosis or heterotopic ossification. Conclusions For patients with complex acetabular fracture involving the posterior column, internal fixation of the anterior acetabulum with a reconstruction plate through the ilioinguinal approach and fixation of the posterior column with antegrade lag screws also through the ilioinguinal approach can result in fine therapeutic effects, because complications like ectopic ossification and sciatic nerve injury related to the Kocher-Langenbeck approach can be prevented. This treatment is particularly suitable for the patients whose condition of the soft tissues at the posterior pelvis is poor.