ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， network pharmacology， molecular docking and cell experiments， the active ingredients， possible targets and molecular mechanisms of Baoxin granules（BXG） regulating ferroptosis in the treatment of heart failure（HF） were explored. MethodsBXG intestinal absorption fluid was prepared by everted gut sac and the chemical composition contained therein were identified by UPLC-Q-TOF-MS. According to the obtained components， the potential targets of BXG were predicted， and the HF-related targets and related genes of ferroptosis were retrieved at the same time， and the intersecting targets were obtained by Venn diagram. In addition， the protein-protein interaction（PPI） network and the component-target network were constructed， and the core components and core targets were obtained by topological analysis. Then Gene Ontology（GO） function and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analysis were performed on the core targets， and molecular docking validation of the key targets and main components was carried out by AutoDockTools 1.5.7. H9c2 cells were used to establish a oxygen-glucose deprivation model， and the protective effect of BXG on cells was investigated by detecting cell viability， cell survival rate and reactive oxygen species（ROS） level. The protein expression levels of signal transducer and activator of transcription 3（STAT3）， phosphorylation（p）-STAT3 and glutathione peroxidase 4（GPX4） were detected by Western blot to clarify the regulatory effect of BXG on ferroptosis. ResultsA total of 61 chemical components in BXG intestinal absorption fluid were identified， and network pharmacology obtained 27 potential targets of BXG for the treatment of HF， as well as 139 signaling pathways. BXG may act on core targets such as STAT3， tumor protein p53（TP53）， epidermal growth factor receptor（EGFR）， JUN and prostaglandin-endoperoxide synthase 2（PTGS2） through core components such as glabrolide and limonin， which in turn intervene in lipid and atherosclerosis， phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt）， endocrine resistance and other signaling pathways to exert therapeutic effects on HF. Molecular docking showed that the docking results of multiple groups of targets and compounds were good. In vitro cell experiments showed that compared with the blank group， the cell viability and survival rate of the model group were significantly decreased， the level of ROS was significantly increased（P<0.01）， the expression levels of STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 proteins were significantly decreased（P<0.05， P<0.01）. Compared with the model group， the cell viability and survival rate of the BXG group were significantly increased， the ROS level was significantly decreased（P<0.01）， the STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 protein levels were significantly increased（P<0.05， P<0.01）. ConclusionBXG may inhibit the occurrence of ferroptosis by up-regulating the expression of STAT3 and GPX4， thus exerting a therapeutic effect on HF， and flavonoids may be the key components of this role.

ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， network pharmacology， molecular docking and cell experiments， the active ingredients， possible targets and molecular mechanisms of Baoxin granules（BXG） regulating ferroptosis in the treatment of heart failure（HF） were explored. MethodsBXG intestinal absorption fluid was prepared by everted gut sac and the chemical composition contained therein were identified by UPLC-Q-TOF-MS. According to the obtained components， the potential targets of BXG were predicted， and the HF-related targets and related genes of ferroptosis were retrieved at the same time， and the intersecting targets were obtained by Venn diagram. In addition， the protein-protein interaction（PPI） network and the component-target network were constructed， and the core components and core targets were obtained by topological analysis. Then Gene Ontology（GO） function and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analysis were performed on the core targets， and molecular docking validation of the key targets and main components was carried out by AutoDockTools 1.5.7. H9c2 cells were used to establish a oxygen-glucose deprivation model， and the protective effect of BXG on cells was investigated by detecting cell viability， cell survival rate and reactive oxygen species（ROS） level. The protein expression levels of signal transducer and activator of transcription 3（STAT3）， phosphorylation（p）-STAT3 and glutathione peroxidase 4（GPX4） were detected by Western blot to clarify the regulatory effect of BXG on ferroptosis. ResultsA total of 61 chemical components in BXG intestinal absorption fluid were identified， and network pharmacology obtained 27 potential targets of BXG for the treatment of HF， as well as 139 signaling pathways. BXG may act on core targets such as STAT3， tumor protein p53（TP53）， epidermal growth factor receptor（EGFR）， JUN and prostaglandin-endoperoxide synthase 2（PTGS2） through core components such as glabrolide and limonin， which in turn intervene in lipid and atherosclerosis， phosphatidylinositol 3-kinase/protein kinase B（PI3K/Akt）， endocrine resistance and other signaling pathways to exert therapeutic effects on HF. Molecular docking showed that the docking results of multiple groups of targets and compounds were good. In vitro cell experiments showed that compared with the blank group， the cell viability and survival rate of the model group were significantly decreased， the level of ROS was significantly increased（P<0.01）， the expression levels of STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 proteins were significantly decreased（P<0.05， P<0.01）. Compared with the model group， the cell viability and survival rate of the BXG group were significantly increased， the ROS level was significantly decreased（P<0.01）， the STAT3， p-STAT3， p-STAT3/STAT3 and GPX4 protein levels were significantly increased（P<0.05， P<0.01）. ConclusionBXG may inhibit the occurrence of ferroptosis by up-regulating the expression of STAT3 and GPX4， thus exerting a therapeutic effect on HF， and flavonoids may be the key components of this role.

Objective: To elucidate the molecular mechanism by which prohibitin 2(PHB2) mediates periodontitis-induced bone tissue inflammation through regulating the nuclear factor kappa B(NF-κB) signaling pathway and its role in irreversible alveolar bone resorption. Methods: Quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR) and immunohistochemistry(IHC) were used to detect the expression differences of inflammatory factors and PHB2 in healthy and inflamed alveolar bone tissues of mice in vivo. In vitro, an inflammatory model was established using lipopolysaccharide(LPS)-induced a mouse calvaria-derived preosteoblastic cell line, subclone E1(MC3T3-E1) cells. Western blot and qRT-PCR were used to clarify the regulatory relationship between PHB2 and inflammatory factors, and immunofluorescence staining was performed to observe changes in PHB2 subcellular localization. PHB2 overexpression plasmids were constructed using molecular cloning, and RNA interference was employed to knock down PHB2 expression to assess its regulatory role in inflammation. Based on RNA-seq data, differential expression analysis based on the negative binomial distribution, version 2(DESeq2) was used for differential expression analysis, and kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment along with gene ontology(GO) functional annotation were performed to identify key signaling pathways and differentially expressed genes. Results: In the mouse periodontitis model, PHB2 expression was significantly upregulated in alveolar bone tissues. In the in vitro inflammatory cell model, PHB2 levels positively correlated with interleukin(IL)-6, IL-1β, and tumor necrosis factor-alpha(TNF-α) levels, and its subcellular localization shifted during inflammation. RNA-seq data and the detection of the level of phosphorylation of p65 protein(p-p65) demonstrated that PHB2 exacerbated inflammatory responses through the NF-κB signaling pathway and was mechanistically linked to upregulation of the upstream chemokine C-X-C motif chemokine ligand 10(CXCL10). Conclusion PHB2 aggravates LPS-induced periodontitis inflammation via the NF-κB signaling pathway, providing new insights into the molecular mechanisms underlying the development of periodontitis.

Objective To explore the prognostic value of sarcopenia in patients undergoing pancreati-coduodenectomy.Method Clinicopathologic data and follow-up information of 116 patients undergoing pancre-aticoduodenectomy at Renmin Hospital of Wuhan University between March 2011 and August 2016 were collected for statistical analysis.Results Among the 116 patients,the prevalence of sarcopenia was 42.2％ (n =49).When compared to the rest of the patients who did not have sarcopenia,the sarcopenia group had longer recovery time [(17.33±6.54) d vs.(13.46±9.32) d,P=0.013] and increased risk of complications (complications in general,59.2％ vs.38.8％,x2 =4.714,P =0.030;Clavien-Dindo ≥ 3:26.5％ vs.10.4％,x2 =5.130,P=0.024).Both the Kaplan-Meier survival analysis (P＜0.05) and the Cox proportional hazard model (overall survival:hazard ratio =2.285,95％ CI =1.521-3.431;recurrence-free survival,hazard ratio =2.167,95％ CI=1.445-3.248) indicated sarcopenia as the risk factor for poorer overall survival and recurrence-free survival.Conclusions Sarcopenia was an independent predictor of poor prognosis for patients undergoing pancreaticoduodenectomy.Patients with sarcopenia had higher risk of developing complications after surgery and lower overall survival rate and recurrence-free survival rate.

Objective: To investigate the prevalence of liver depression and spleen deficiency syndrome in sub-health status and the demographic data of liver depression and spleen deficiency syndrome in sub-health status. Methods: The clinical epidemiological survey was conducted in the routine physical examination population using the "Sub-health State TCM Syndrome Questionnaire (3 Edition)". Descriptive analysis was used to analyze the relevant data of sub-health status of liver depression and spleen deficiency syndrome. Results: The incidence of sub-health state of liver depression and spleen deficiency syndrome was 7.16％. The sub-health state of liver depression and spleen deficiency syndrome was characterized by a higher proportion of females than males. The incidence of liver depression and spleen deficiency syndrome was higher in all groups over 19 years of age. The highest proportion occurred in colleges and universities, and the highest proportion occurred in divorced persons in marital status. The occupational status was the highest among persons in charge of enterprises and institutions, administrative personnel, professional technicians, and teachers with administrative personnel.

Objective: To observe the changes of Notch signaling pathway related molecule expression in hippocampus of rats with stagnation of liver qi and spleen deficiency syndrome and to explore the regulatory effect of Xiaoyaosan. Methods: Male SD rats were randomly divided into four groups: normal group, model group, model+xiayaosan group and model+fluoxetine group, each group had 12 rats. The stagnation of liver qi and spleen deficiency model was established by chronic immobilization stress for 21 days. Detection of Nissl bodies by Nissl staining, expression of NICD, Hes1, Hes5 and Jad1 were detected by Fluorescent quantitative PCR and Western-blot method. Results: Compared with the normal group, the number of Nissl bodies in the model group decreased significantly (P ＜ 0.01), which showed that Xiaoyaosan and fluoxetine could reverse it (P ＜ 0.01). The expression of Notch signaling pathway-related protein in model group was significantly lower than that in normal group (P ＜ 0.01), which showed that Xiaoyaosan and fluoxetine could reverse the expression of NICD, Hes5 and Jag1 protein (P ＜ 0.05, P ＜ 0.01). Compared with the normal group, the expression of each gene in the model group decreased (P ＜ 0.01), which showed that Xiaoyaosan and fluoxetine could reverse the expression of NICD, Hes1 and Hes5 mRNA (P ＜ 0.05, P ＜ 0.01). Conclusion: The hippocampal neurons of rats with stagnation of liver qi and spleen deficiency syndrome were damaged and the Nissl bodies were reduced, the expression of each protein and gene in Notch signaling pathway decreased. Xiaoyaosan may play a therapeutic role by regulating the expression level of hippocampal related molecules to protect neurons.

Objective To investigate the changes and clinical significance of neck-shaft angle in the treatment of proximal humerus fractures with locking plate.Methods A retrospective case-control study was done on 214 cases of proximal humerus fractures treated with locking plate in our hospital from January 2008 to June 2016.According to the presence of medial bone defects,the cases were divided into medial bone defect group (85 cases) and non-medial bone defect group (129 cases).In medial bone defect group,there were 35 males and 50 females,with an average age of 63.3 years old.In this group,the cases of Neer three-part fractures and Neer four-part fractures were 47 and 38,respectively.In non-medial bone defect group,there were 51 males and 78 females,with an average age of 65.9 years old.In this group,the cases of Neer three-part fractures and Neer four-part fractures were 57 and 72,respectively.The preoperative humeral neck-shaft angle of contralateral humerus,and the humeral neck-shaft angle with its changes on the second postoperative day as well as 1,3,6 and 12 months after surgery were evaluated by the X-ray film.The Constant scores of the shoulder function were also assessed.Results The follow-up time for the patients ranged from 12 to 42 months (mean 24.2 months).In medial bone defect group,the preoperative neck-shaft angle of contralateral humerus,as well as neck-shaft angles of fractured side at five follow-up visits were (133.4±4.0) °,(134.7 ±11.5) °,(120.8±4.0) °,(118.5 ±3.5) °,(117.9 ±10.4) °and (114.7 ±4.4)°,respectively.While in the other group,the corresponding parameters were (134.0±2.4)°,(133.8± 17.1)°,(135.6± 2.9)°,(132.7± 2.8) °,(131.5 ± 13.5) °,(135.2 ± 2.8) °,respectively.There were significant differences between the two groups of the neck-shaft angle at 1,3,6 and 12 months (P ＜ 0.05).In the medial bone defect group,there were no significant differences between neck-shaft angle of contralateral humerus and the second postoperative day (P ＞0.05),but both of them are significantly greater than others(P ＜0.05).In the other group,except for the neck-shaft angles of postoperative 1 month and 6 months (P ＜ 0.05),there was no statistically significant difference between the angles in different time points(P ＞ 0.05).The mean Constant scores of medial bone defect group and no medial bone defect group were (69.3 ± 14.6) and (75.2 ± 12.6),respectively.Conclusions After the recovery of neck shaft angle,there is still a need of attention to the losses of neck shaft angle after surgery.Neck-shaft angle could be a reference for the treatment strategy of proximal humerus fractures with locking plate,which could guide the anatomical reduction as well as be used to evaluate the effects after surgery.It could also be a reference for function exercise.Moreover,the losses of neck shaft angle will be influenced by supporting from internal side,which could have effects on the function recovery after surgery.

Objective To investigate the association between different number of inferomedial screws (0-3 screws) and their efficacy in locking plate of proximal humerus fractures.Methods Data of 90 patients who had undergone locking plate for proximal humerus fracture between January 2007 and December 2012 were retrospectively analyzed.They were divided into four groups according to the number of inferomedial screws.In the NS (no-screw support) group,36 patients did not have inferomedial screws for mechanical support of the inferomedial region of the proximal humerus; in the OS (one-screw support) group,23 patients had the reconstruction by insertion of only one inferomedial screw; there were 19 patients in the TS (two-screw support) group and 12 in the ThS (three-screw support) group.There were no significant differences among 4 groups in sex,fracture type and age.The Constant scores of the shoulder function,changes in humeral head height at the latest follow-up,incidence of complications and time for fracture healing among the 4 groups were collected.Results The follow-ups for the patients ranged from 12 to 56 months (mean,21.4 months).At the latest follow-up,the ThS,TS,OS and NS groups had a mean Constant score of 76.7 (SD,11.6),74.1 (SD,7.4),66.8 (SD,10.7),65.8 (SD,10.2),respectively; the mean change in humeral head height were 1.4 (SD,1.0) mm,2.9 (SD,1.2) mm,5.3(SD,3.2) mm and 6.7(SD,3.3) mm,respectively.There were no significant differences between the ThS and TS groups in Constant score,and Constant scores in ThS and TS groups were significantly higher than those in OS and NS groups.However,the loss in humeral head height in the ThS group was significantly less than those in the TS,OS and NS groups,and the loss in humeral head height in TS group was significantly less than those in the OS and NS groups.There were no significant differences among 4 groups in time for fracture healing and incidence of complications.Conclusion Locking plate in general do not appear to be a panacea for proximal humerus fractures especially in cases of medial comminution or bone defects because of the malreduction of the medial cortical bone.However,medial support with two or three inferomedial screws being meticulously placed in the inferomedial region may enhance the mechanical stability of medial column support and allow for better maintenance of reduction.

Objective To evaluate correlation of radial head fracture with forearm interosseous membrane (IOM) injury.Methods Twenty-six patients with radial head fractures were studied prospectively between September 2007 and June 2010.There were 15 men and 11 women,with an average age of 37.6years (range,21-53).According to the Mason classification,there were 7 cases of type Ⅰ,9 cases of type Ⅱ,10 cases of type ⅢL All patients were subjected to forearm X-ray,CT scans and the MR within a week.Clinical and radiographic data of all the patients were collected.Spearman rank correlation statistical analysis was used to analyze the correlation between the radial head fracture and the IOM injury.Results The radial head fractures and IOM injury were directly related.The IOM injury was noted in all type of radial head fracture.The more severity radial head fracture had,the more IOM injury happened.In Mason Ⅰ-Ⅲ fractures,IOM injury was found in 2,4 and 7 cases respectively.The different degree of radial head fracture caused different effects on IOM injury.The severity of radial head fracture was correlated with damage degree of IOM.In Mason type Ⅰ and type Ⅱ fractures,the IOM injury were just partial disruption with distal part of the IOM and did not reach the biomechanically essential central band.In type Ⅲ fractures,central band disruption was found in 3 cases.Conclusion Mason Ⅰ-Ⅲ radial head fractures are associated with forearm IOM injury.There was a positive correlation between radial head fractures and IOM injury.If IOM lesions are suspected,magnetic resonance imaging should be performed,especially Mason Ⅲ fractures.

Objective To improve the accuracy of X-ray diagnosis of the deltoid ligament injury by evaluating the influence of plantar flexion on the diagnosis.Methods Twenty-four patients with ankle supination-extemal rotation fracture which was distal fibula fracture but no medial malleolus fracture were treated from February 2010 to December 2010.All the patients were taken Mortise radiograph with the ankle in four positions of plantar flexion(0°,15°,30° and 45°)applied with rotational stress and the deltoid ligament injury was identified with MRI.The medical clear space(MCS)and the superior clear space(SCS)on each radiograph were measured.The results were tested by a one-way analysis of variance model(ANOVA),based on the following criteria:(1)the MCS≥ 4 mm and MCS ＞ SCS,or when MCS≥5 mm and MCS ＞SCS.The MRI examination was taken as the gold standard to compare the influence of X-ray diagnosis of the deltoid ligament injury with different plantar flexions and different standards.Results The average MCS in four positions of plantar flexion(0°,15°,30°,and 45 °)with rotational stress were(4.10 ±0.79)mm,(4.55 ±0.72)mm,(4.99 ±0.56)mm and(5.71 ±0.86)ram,with statistical difference(P ＜0.05).The average SCS in four positions of plantar flexion(0°,15°,30°,and 45°)were(3.56 ±0.41)mm,(3.50 ±0.43)mm,(3.71 +0.44)mm and(3.93 ±0.51)mm,with no statistical difference in the four groups(P ＞0.05).With the MSC≥4 mm and MSC ＞ SCS as the diagnostic criteria,the prevalence of false-positive findings of deltoid injury based on the ratio with the four positions of plantar flexion(0°,15°,30° and 45°)were 50.0％,66.7％,88.9％ and 94.4％ respectively.With the MSC≥5 mm and MSC ＞SCS as the diagnostic criteria,the prevalence of false-positive findings of deltoid injury based on the ratio with the four positions of plantar flexion(0°,15°,30° and 45°)were 5.6％,11.1％,38.9％ and 77.8％ respectively.Condusions Different plantar flexion angle is an important factor for X-ray diagnosis of deltoid ligament injury of the ankle joint.The prevalence of false-positive findings of deltoid injury increases with the increase of plantar flexion.