Objective To investigate the preventive effect of schisandrin (SCH) on fetal neural tube defects (NTDs) of mice and its mechanism. Methods C57BL/6 mice were mated with female and male at a ratio of 2:1. Pregnant female mice with vaginal plug after mating were randomly divided into control group, model group, SCH group, and folic acid group, with 9 mice in each group. The NTDs fetal mice model was induced by intraperitoneal injection of all-trans retinoic acid (atRA) (7.5 mg/kg) on embryonic day 7.5 (E 7.5 d). During E 0.5 d-E 11.5 d, pregnant rats in folic acid group were given folic acid [61.0 μg/(kg·d)] by gavage once a day, and pregnant rats in SCH group were given SCH [8.0 mg/(kg·d)] by gavage once a day. Fetal mice were removed by cesarean section on E 11.5 d. PC12 cells were divided into control group, model group and SCH group. PC12 cells were treated with atRA (20 μmol/L) for 12 hours to establish cell damage model in model group, and treated with SCH (2.5 μmol/L) for 24 hours in SCH group. Fetuses were identified NTDs by stereoscopic microscopy. HE staining was used to observe the closure of the neural tube. The expression levels of p-PI3K, Akt and p-Akt molecules in PI3K/Akt signaling pathway were detected by Western Blotting. Results Compared with control group, the incidence of NTDs was significantly increased in mice of model group (P<0.01); compared with model group, the incidence of NTDs was decreased in folic acid group and SCH group (P<0.01); compared with folic acid group, SCH group had a lower incidence of NTDs (P<0.01). Western Blotting results showed that compared with control group, the expression of p-PI3K and p-Akt protein in fetal tissues of model group was significantly decreased (P<0.01, P<0.05); compared with model group, there was no significant difference in expression of p-PI3K and p-Akt in fetal tissues of folic acid group (P>0.05), while the expression of p-PI3K and p-Akt protein in SCH group was significantly higher (P<0.05). Compared with control group, PC12 cells in model group showed lower expression levels of p-PI3K and p-Akt (P<0.05); compared with model group, PC12 cells in SCH group showed higher expression levels of p-PI3K and p-Akt (P<0.05). Conclusions SCH can reduce the incidence of atRA-induced NTDs in fetal mice, and its preventive effect is better than folic acid, which may be related to the activation of the PI3K/Akt signaling pathway.

Objective Establishing animal model of Vibrio vulnificus (V. vulnificus) early infection via respiratory tract, to observe the survival, pathological changes of target organs and the expression changes of inflammatory cytokines in model mice, and to provide references for the relevant prevention and treatment strategies. Methods A total of 24 healthy female BALB/c mice (8 to 10 weeks old) were randomly divided into 4 groups (6 mice per group), including control group and three infection groups (low, medium and high concentration of infection, respectively), based on which the V. vulnificus infection models were established by intranasal administration. The survival statistics, status of defecation, hair and respiration as well as mental state of mice were monitored during 0-12 h early infection progress. Hematoxylin-eosin (HE) staining was performed on the important organs of mice to analyze the pathological manifestations, and immunohistochemical staining was used to detect the expression of interleukin (IL)-6, IL-10, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in the target organs. Mouse inflammation panel combined with flow cytometry was used to further detect the changes of multiple inflammatory and anti-inflammatory cytokines in serum. Results V. vulnificus infection via respiratory tract caused mice deterioration in survival rate, defecation, hair status, respiratory tract and mental state within 12 h. HE pathological analysis showed inflammatory cell infiltration and cell necrosis in ileum, lung, liver and spleen of mice in different infection groups, and the injury was much severer in high concentration of infection group. Immunohistochemical results reflected the positive expression rates of IL-6, IL-10, IFN-γ and TNF-α in ileum, lung, liver and spleen of mice in medium and high concentration of infection groups were significantly higher than those in control group (P<0.05). The results of mouse inflammation panel and flow cytometry showed that compared with the control group, the expression levels of TNF-α and monocyte chemotactic protein (MCP)-1 in all three infection groups were significantly increased (P<0.05), and the expression level of IL-27 significantly decreased (P<0.05); the expression levels of IL-1β, IL-6, IL-17A and granulocyte-macrophage colony-stimulating factor (GM-CSF) were significantly increased in medium and high concentration of infection groups (P<0.05); the expression levels of IL-1α, IL-23, IL-12p70 and IFN-γ were significantly increased in high concentration of infection group (P<0.05). Conclusions V. vulnificus infection progressed rapidly through respiratory tract in mice, and lung, intestine, liver and spleen were major target organs. Accompanied by increased secretion of multiple serum inflammatory cytokines, V. vulnificus infection through respiratory tract might further causes severe inflammatory reaction in hosts.

Objective To investigate the EMG characteristics and clinical application value of surface EMG in cervical muscle group of children with congenital muscular torticollis (CMT). Methods A retrospective analysis was performed on 478 children diagnosed as CMT from January 2018 to August 2022 in Rehabilitation Center of the Children's Hospital Affiliated to Chongqing Medical University. The torticollis side of the same case was included in observation group and the healthy side was included in control group. Surface EMG was used to record the electromyography root-mean-square values of bilateral sternocleidomastoid muscles, platysma muscles and upper trapezius muscles in the rest and different exercise states. The difference of root-mean-square values between the affected side and the healthy side muscle groups at rest and under different exercise states and the diagnostic value were compared. Spearman correlation analysis was used to evaluate the relationship between the severity of congenital muscular torticollis on the affected side and the root mean square value of each cervical muscle group. Results The root-mean-square value of sternocleidomastoid muscle on the affected side of CMT children was higher than that on the healthy side during relaxation and lower than that on the healthy side during contraction, and the differences were statistically significant (P<0.05), the abnormal root-mean-square value of the affected side in the resting position and in all motion state has higher diagnostic value. The root-mean-square value of platysma muscle on the affected side was higher than that on the healthy side when it was in the resting position and the reverse neck rotation relaxed state (P<0.05), and lower than that on the healthy side when it was in the same neck rotation contraction state (P<0.05), abnormal values detected in resting sitting position and elevation of head have higher value. Root-mean-square value of upper trapezius muscle on the affected side was higher than that on the healthy side in the relaxed state (P<0.05). The abnormal values in sitting position, turning neck and bending head have high diagnostic value. The severity of CMT was positively correlated with the root-mean-square values of sternocleidomastoid muscle, platysma muscle, and upper trapezius muscle (r was 0.721, 0.612, 0.655, P<0.001). Conclusion sEMG could be used as a good method for the clinical diagnosis and evaluation of congenital muscular torticollis in children.

Objective To explore the pattern of early expression and secretion of tissue factor (TF) in vascular endothelial cells induced by heat stress. Methods Thirty SPF-rated C57BL/6 male mice were randomly divided into five groups: the control group and groups of indicated recovery time, including 0, 3, 6, and 9 h in room temperature after heat stress (n=6). Mice in the heat stress groups were exposed to an animal incubator to reach 42.5 ℃ for core body temperature for heat stroke. We analyzed the histopathological changes in the liver, lung, and kidney tissues with HE staining. We measured the TF mRNA in mice tissues by RT-qPCR and the plasma concentration of TF in mice with a commercial ELISA kit. Human umbilical vein endothelial cells (HUVECs) were placed in a culture incubator to build an in vitro heat stress model. HUVECs were divided into five groups, including a control group and groups of indicated recovery time, including 0, 3, 6, and 9 h after heat stress. We quantified the expression of TF mRNA and protein in HUVEC cells by RT-qPCR, Western blotting, and immunofluorescence and measured the secreted TF with a commercial ELISA kit. Results No significant pathological injury was observed in the tissues of the control group. Mice treated with heat stress had various degrees of structural injuries and hemorrhagic and inflammatory changes in multiple tissues. Compared to control group, the expression of TF mRNA significantly increased in the kidney of heat stress-treated mice with 0 and 3 h recovery time (1.719±0.018, 1.241±0.178 vs. 1.000±0.063), the lung with 3 h recovery time (2.444±0.511 vs. 1.000±0.106) and the liver with 6 h recovery time (7.312±0.618 vs. 1.000±0.147) (P<0.05). The concentration of TF in plasma also sustainedly elevated in mice with 0, 3, 6, and 9 h recovery time after heat stress as compared to control group [(132.426±17.920) pg/ml, (119.400±10.267) pg/ml, (107.374±13.495) pg/ml, (163.767±22.810) pg/ml vs. (75.479±13.831) pg/ml, respectively, P<0.01]. The expression levels of TF mRNA were higher in heat stress HUVECs with 6 h and 9 h recovery time than the control cells (1.905±0.354, 2.564±0.297 vs. 1.000±0.097, P<0.01). Secreted TF in the supernatant from HUVECs treated with heat stress and different recovery time also increased significantly [(36.309±4.101) pg/ml, (38.425±5.484) pg/ml, (41.655±4.380) pg/ml, (43.586±4.718) pg/ml vs. (14.996±0.254) pg/ml, P<0.01]. Conclusion Heat stress increased early expression and secretion of TF in vascular endothelial cells. Vascular endothelial cells may be a main source of circulating TF in heat stroke.

Objective To report 5 cases of amyloid arthropathy with joint flexion contracture as the first manifestation, and perform a literature review to improve the understanding of the disease. Methods Retrospective analysis of the clinical manifestations, laboratory and imaging results clinical data of patients with immunoglobulin light chain (AL) amyloidosis related amyloid arthropathy with joints flexion contracture as the first manifestation at the Department of Rheumatology and Immunology, the First Medical Center of People's Liberation Army General Hospital from January 2012 to February 2022. Meanwhile searching the database (CNKI, Wanfang Data, PubMed), excluding myeloma-associated amyloid arthropathy, and analyzing the literature results comprehensively. Results There were a total of 5 patients with amyloid arthropathy with joint flexion contracture as the initial manifestation in this center, including 4 males and 1 female. By February 2022 (searching CNKI, Wanfang Data and PubMed), two eligible patients were selected, all males. A total of 7 patients were included with male-to-female ratio of 6:1. The median age was 30-year-old (range from 19 to 73). Spine and limb joints were involved, especially finger joints (7 cases, 100%). Soft tissue was involved in 5 patients (71.4%), carpal tunnel syndrome in 4 patients (57.1%), giant tongue in 3 patients (42.9%), shoulder pad sign in 2 patients (28.6%), muscle swelling in 2 patients (28.6%). Five patients were misdiagnosed as rheumatoid arthritis (RA) and 1 patient was misdiagnosed as spondyloarthritis (SpA). The median time of delayed diagnosis was 24 months (2-204 months). The laboratory results showed that 3 patients (42.9%) were positive for immunoelectrophoresis, all of which were λ light chain. Only 1 patient was positive for HLA-B27, while the other patients were negative for autoantibodies and rheumatoid factors. Only 1 patient had elevated C reactive protein (CRP) and erythrocyte sedimentation rate (ESR). Ultrasound examination of the lesion site of 5 patients (71.4%) showed hypoechoic area around the joint, thickening of ligaments or periarticular tenosynovitis. Five patients (71.4%) underwent X-ray examination, of which 4 patients had no bone destruction, 1 patient had soft tissue swelling with bone erosion and subchondral cyst. MRI was performed in 5 patients (71.4%), in which low signal on T₁ weighted image, medium signal on T₂ weighted image and edema signal on peripheral tendon sheath were seen in 1 patient. One patient (14.3%) underwent emission computed tomography (ECT) without abnormal uptake. Conclusion Joint flexion contracture may be the first sign of amyloid arthropathy. Vigilance for amyloid arthropathy is warranted when there is associated soft tissue involvement, negative autoantibodies, and imaging showing no joint destruction but surrounding soft tissue abnormalities.

Objective To explore the protective effects and mechanisms of cysteinyl leukotriene 2 (CysLT₂) receptor antagonist HAMI3379 on cerebral ischemia injury in rats. Methods 30 male SD rats were randomly divided into sham operation group, model group, and HAMI3379 group, with 10 rats in each group. The rats of model group were subjected to middle cerebral artery occlusion (MCAO) to construct the cerebral ischemia injury model, while HAMI3379 group received intraperitoneal injection of HAMI3379 (0.2 mg/kg) before and after MCAO 30 min. The rats after cerebral ischemia injury were scored for neurological symptoms. The infarction volume of rats was observed by TTC staining, the activation marker Iba1 of microglia was detected by immunofluorescence staining, the mRNA level of M1/M2 polarized phenotype molecules of microglia was detected by Real-time PCR, the number of neurons was observed by NeuN staining, and neuronal degeneration was observed by Fluoro-Jade B staining. Western blotting assay was used to detect the expression of CysLT₂ protein and nuclear factors κB-related protein Cα (PKCα), IκBα, p65 and p50 proteins in brain tissue. Results Compared with sham operation group, the neurological symptom score and cerebral infarction volume of model group were significantly increased (P<0.05). Compared with model group, the neurological symptom score and cerebral infarction volume of HAMI3379 group were significantly decreased (P<0.05). The results of immunofluorescence staining showed that the expression of microglia activation marker Iba1 was increased in brain tissue of rats after cerebral ischemia injury (P<0.05). Compared with sham operation group, mRNA expression of M1 polarized molecules (CD86, IL-1β, TNF-α) and M2 polarized molecules (CD206, TGF-β, IL-10) were significantly increased in the ischemic central brain tissue of model group (P<0.05). Compared with model group, the expression of M1 polarized molecules in HAMI3379 group was significantly downregulated (P<0.05), while the expression of M2 polarized molecules was significantly upregulated (P<0.05). Compared with sham operation group, the expressions of PKCα, IκBα, p65 and p50 in brain tissue of model group were significantly up-regulated (P<0.05); compared with model group, the expressions of PKCα, IκBα, p65, and p50 in HAMI3379 group were significantly down-regulated (P<0.05). The NeuN staining results showed that the number of neurons in the brain tissue of model group was decreased when compared with sham operation group (P<0.05), while the number of degenerated neurons was increased (P<0.05). Compared with model group, the number of neurons in HAMI3379 group was increased (P<0.05), while the number of degenerated neurons was decreased (P<0.05). Conclusions CysLT₂ receptor antagonist HAMI3379 may regulate PKCα/IκBα/NF-κB signaling pathway, inhibits M1 polarization activation of microglia and promotes its transition to M2 polarization, inhibits neuronal degeneration, and plays a neuroprotective role.

The prothrombin complex concentrate (PCC) is a blood product that mainly contains vitamin K-dependent coagulation factors. PCC has been used clinically for more than 70 years, and its safety has been significantly improved. It has been widely used in the treatment of coagulation disorders or severe bleeding in critically ill patients. If used improperly, it may weaken the hemostatic effect or increase the risk of thrombosis. In order to standardize the rational use of PCC, Chinese Society of Thrombosis, Hemostasis and Critical Care, Chinese Medicine Education Association, People's Liberation Army Professional Committee of Critical Care Medicine, Chinese Society of Thrombosis and Hemostasis, Chinese Research Hospital Association have organized the formulation of this expert consensus, including three parts: definition, indications, and monitoring and evaluation of efficacy, with a total of 10 suggestions, aiming to help clinicians reasonably use PCC and improve the treatment in critical illness.

Tracheoesophageal fistulas (TEF) is a kind of pathological fistula between trachea or bronchus and esophagus, including tracheoesophageal fistula and bronchoesophageal fistula. In clinical practice, tracheoesophageal-fistula is more often seen. There are many pathogenesis of TEF, which could cause serious complications. It is a disease which has serious impact on the quality of life, difficult to treat and high mortality. There are many methods to treat TEF, but the therapeutic effect is poor. There is an urgent need for new treatment methods to TEF. The detection of nasogastric tube retention by chest and abdominal X-ray is the gold standard for diagnosis. The main treatment methods include surgical treatment, stent implantation, local biological glue blocking, stem cell therapy and so on. In order to provide reference for relevant clinical diagnosis and treatment of TEF, this article reviews the main research progress in the diagnosis and treatment of TEF in recent years.

Objective To investigate the improvement effect and potential mechanism of m⁶A recognition protein YTHDF1 on Duchenne muscular dystrophy (DMD). Methods (1) We collected muscle tissue and peripheral blood from DMD patients (38 confirmed patients with surgery at Xi'an Children's Hospital from September 2016 to June 2017, DMD group) and non-DMD patients with orthopedic surgery (33 age-matched patients, 2-3 years old, control group). The expression levels of YTHDF1 and dystrophin in the muscle tissues were quantified by RT-qPCR and Western blotting. The serum creatine phosphokinase (CPK) content was determined using ELISA. (2) Take SkMC from skeletal muscle myoblasts and set up empty vector group (transfected with scrambled negative control), AAV-YTHDF1 group transfected with YTHDF1 overexpression vector and scrambled group (transfected with NC siRNA) and si-YTHDF1 group (transfected with YTHDF1 siRNA). Cell proliferation was analyzed using an EdU assay. The expressions of dystrophin, myogenin (MyoG), and myosin heavy chain (MHC) proteins were measured by Western blotting. A RIP assay was used to verify the interaction between YTHDF1 and YES-associated protein 1 (YAP1) mRNA and investigate the modification level of the m⁶A site on YAP1 mRNA. (3) Thirty Mdx mice were randomly divided into: the empty vector group (n=15, intraperitoneal injection of empty adenoviral empty vector) and the AAV-YTHDF1 group (n=15, intraperitoneal injection of YTHDF1 overexpression adenovirus vector). We profiled the body weights, muscle and organ wet weights, fiber diameter, and fiber type of the mice. The protein levels of YTHDF1 and dystrophin were detected using Western blotting. We used HE staining to observe the pathological changes in the gastrocnemius and quadriceps femoris muscles. Results (1) Compared with control group, the expression of YTHDF1 and dystrophin was significantly lower in DMD patients (P<0.01), and the serum CPK content was significantly increased (P<0.0001). (2) Compared with empty vector group, the EdU positive rate and the expression of dystrophin, MyoG, and MHC proteins significantly increased (P<0.01). In addition, overexpression of YTHDF1 prolonged the half-life and improved the stability of YAP1 mRNA. Compared with Scramble group, the EdU-positive rate and the expression of dystrophin, MyoG, and MHC proteins significantly decreased (P<0.05 or P<0.01). (3) Compared with empty vector group, the mice in AAV-YTHDF1 group developed more rapidly with higher muscle mass in the gastrocnemius muscle, quadriceps muscle, and triceps (P<0.05). We observed no difference in the weights of inguinal, gonadal, or retroperitoneal fat pads and a significant increase in the fiber areas of gastrocnemius and quadriceps femoris (P>0.05). Conclusion YTHDF1 regulates the stability of YAP1 mRNA by recognizing the m⁶A modification of YAP1 mRNA and thus promotes YAP1/dystrophin-mediated myocyte proliferation to inhibit DMD progression.

Objective To investigate the efficacy and safety of stereotactic body radiotherapy (SBRT) combined with camrelizumab and apatinib in treatment of advanced hepatocellular carcinoma (HCC). Methods The clinical data were retrospectively analyzed of 85 patients with advanced HCC treated in the Fifth Affiliated Hospital of Zhengzhou University and People's Hospital of Zhengzhou from January 2019 to September 2021. They were divided into observation group (n=31, SBRT combined with camrelizumab and apatinib) and control group (n=54, treated with camrelizumab and apatinib) according to whether they received SBRT. The propensity score matching (PSM) was used to balance the influence of confounding factors. The objective remission rate (ORR) and disease control rate (DCR) were compared between the two groups. The 6-month overall survival rate, 1-year overall survival rate and progression-free survival (PFS) were compared between the two groups by Kaplan-Meier method. The safety of the two groups was evaluated by Common Terminology Criteria for Adverse Events (CTCAE) version 5.0. Results Before PSM, there were significant differences in age (P=0.043), number of extrahepatic metastasis (P=0.028), and previous surgical treatment (P=0.038) between the two groups. After PSM, there was no significant difference in baseline characteristics between the two groups (P>0.05). After PSM, 27 cases were included in each groups, and three months after treatment, the ORR in observation group and control group were 66.7% and 29.6%, respectively, showed difference with statistically significant (P=0.006); and the DCR in the both groups were 96.3% and 85.2% respectively, showed no statistically significant difference (P=0.175). There were statistical differences in 6-month overall survival rate (96.3% vs. 80.9%, P=0.001), 1-year overall survival rate (75.0% vs. 61.4%, P=0.034) and median PFS (8 months vs. 5 months, P=0.003) between the observation group and control group. Multi-factor Cox regression analysis showed that baseline alpha-fetoprotein (AFP) ≥400 ng/ml was an independent risk factor for affecting the survival of patients with advanced HCC (HR>1, P<0.05), while the triple therapy and previous targeted drugs therapy were the protective factors for the survival of patients with advanced HCC (HR<1, P<0.05). In the observation group, 4 patients had grade 3 adverse reactions, and the common adverse reactions were dyspepsia (14.8%). One case of grade 3 adverse reactions occurred in control group, and there was no statistically significant difference in the incidence of adverse reactions between the two groups (P=0.639). Conclusion SBRT combined with camrelizumab and apatinib is a safe and effective treatment for advanced HCC with significant clinical effect and controllable adverse reactions.