Objective To investigate the therapeutic effects and mechanisms of Aurantii Fructus Immaturus(AFI)on slow-transit constipation(STC)in mice.Methods A STC model was established via intragastric administration of Loperamide Hydrochloride.Successfully modeled mice were randomized into a model group,low-and high-dose AFI groups,a high-dose AFI+Masitinib[tyrosine kinase(c-Kit)inhibitor]group,additionally,a normal group was set up.After intervention,intestinal transit rate,6-hour fecal pellet number,fecal water content,and colonic histomorphology(hematoxylin-eosin staining)were assessed.Enzyme-linked immunosorbent assay(ELISA)was used to measure levels of 5-hydroxytryptamine(5-HT),vasoactive intestinal polypeptide(VIP),substance P(SP),nitric oxide(NO),and nitric oxide synthase(NOS)activity in colon tissue.Quantitative real-time PCR(qRT-PCR)was used to evaluate mRNA expression of stem cell factor(SCF)and c-Kit in colon tissue,Western Blot was used to analyze relative protein expression of aquaporin 3(AQP3),aquaporin 8(AQP8),SCF,and c-Kit in colon tissue.Results Compared with the normal group,there was devere colonic damage observed in the colon tissue of the mice in the model group,the fecal pellet number,fecal water content,intestinal transit rate,colon tissue SP content,c-Kit and SCF gene and protein expression levels were reduced.The contents of 5-HT,VIP and NO,NOS activity and the protein expression levels of AQP3 and AQP8 in colon tissue were increased,and the differences were statistically significant(P＜0.05).Compared with the model group,the colonic injury of mice in the low-and high-does AFI groups showed obvious improvement,the fecal pellet number,fecal water content,intestinal transit rate,colon tissue SP content,c-Kit and SCF gene and protein expression levels were increased,the contents of 5-HT,VIP and NO,NOS activity and the protein expression levels of AQP3 and AQP8 in colon tissue were decreased(P＜0.05).Masitinib partially reversed the laxative effects of AFI(P＜0.05).Conclusion AFI enhances intestinal motility,alleviates colonic injury,and improves defecation in STC mice,potentially via activation of the SCF/c-Kit pathway.

Peri-implant diseases have a relatively high incidence,and immune-inflammatory re-sponses are closely associated with peri-implant mucositis and peri-implantitis.The complement sys-tem is a vital component of the immune system.Its activation not only regulates the initiation and pro-gression of inflammatory responses but also plays a crucial role in bone remodeling.This review sum-marizes the roles of the core complement components C3 and C5 in various stages of bone remodeling after implant placement,as well as in peri-implant mucositis and peri-implantitis.The aim of the pa-per was to provide novel insights for the prevention and treatment of peri-implantitis through targeted regulation of complement protein levels.

Objective Tto evaluate the effect of low intensity pulsed ultrasound(LIPUS)on the migration and phagocytosis ability of lipopolysaccharide(LPS)-induced RAW264.7 macrophages and on macrophage behavior under inflammation.Methods An in vitro activated RAW264.7 macrophage model was developed using LPS.Cell viability was assessed using a CCK-8 assay to explore the effect of LIPUS on activated and inactivated macrophages.Wound healing assays were employed to measure the effect of LIPUS on macrophage migration,and the Transwell assay was employed when LPS was used as a chemoattractant.Phagocytosis ability was examined using confocal microscopy and flow cytometry by observing the FITC fluorescence signal of internalized pHrodo Green E.coli BioParticles Conjugate.Results An activated RAW264.7 macrophage model was successfully developed using 100 ng/mL LPS.LIPUS inhibited the migration of inactivated macrophages into scratch areas as well as guided cell migration.However,cell viability and phagocytosis remained unchanged.Conclusion LIPUS may inhibit RAW264.7 migration but not affect the phagocytosis ability of the macrophages.

Background and objective Invasive mucinous adenocarcinoma(IMA)was a rare and specific type of lung adenocarcinoma,which was often characterized by fewer lymphatic metastases.Therefore,it was difficult to evaluate the prognosis of these tumors based on the existing tumor-node-metastasis(TNM)staging.So,this study aimed to develop Nomo-grams to predict outcomes of patients with pathologic N0 in resected IMA.Methods According to the inclusion criteria and exclusion criteria,IMA patients with pathologic N0 in The Affiliated Lihuili Hospital of Ningbo University(training cohort,n=78)and Ningbo No.2 Hospital(validation cohort,n=66)were reviewed between July 2012 and May 2017.The prognostic value of the clinicopathological features in the training cohort was analyzed and prognostic prediction models were established,and the performances of models were evaluated.Finally,the validation cohort data was put in for external validation.Results Univariate analysis showed that pneumonic type,larger tumor size,mixed mucinous/non-mucinous component,and higher overall stage were significant influence factors of 5-year progression-free survival(PFS)and overall survival(OS).Multivariate analysis further indicated that type of imaging,tumor size,mucinous component were the independent prognostic factors for poor 5-year PFS and OS.Moreover,the 5-year PFS and OS rates were 62.82％and 75.64％,respectively.In subgroups,the sur-vival analysis also showed that the pneumonic type and mixed mucinous/non-mucinous patients had significantly poorer 5-year PFS and OS compared with solitary type and pure mucinous patients,respectively.The C-index of Nomograms with 5-year PFS and OS were 0.815(95％CI:0.741-0.889)and 0.767(95％CI:0.669-0.865).The calibration curve and decision curve analysis(DCA)of both models showed good predictive performances in both cohorts.Conclusion The Nomograms based on clinicopathological characteristics in a certain extent,can be used as an effective prognostic tool for patients with pathologic N0 after IMA resection.

OBJECTIVES: To explore the neuroprotective mechanism of electroacupuncture at the acupoints Baihui and Shenting in rats with cerebral ischemia-reperfusion (IR) injury. METHODS: Forty-eight male SD rats were equally randomized into sham operation group, cerebral IR model group, acupoint electroacupuncture group and non-acupoint acupuncture group. In the latter 3 groups, cerebral focal ischemic injury was induced using the Longa method; in the two electroacupuncture groups, electroacupuncture was performed either at the acupoints Baihui and Shenting or at non-acupoint sites for 7 days. The changes in neurological deficit scores, cerebral infarction volume, learning and memory function, pathologies in hippocampal CA1 area, neuronal and synaptic ultrastructures, and synaptic density of the rats were observed, and serum GABA level and mRNA and protein expressions of GABA_AR α1, CaMK II, SYN1 and PSD-95 in the hippocampal tissue were detected. RESULTS: Compared with those in cerebral IR model group, the rats receiving electroacupuncture at the acupoints, but not those with electroacupuncture at the non-acupoints, showed significantly decreased neurological deficit scores and cerebral infarction volume with shortened escape latency and increased platform crossings. Electroacupuncture at the acupoints significantly increased neuronal cell number, decreased the width of the synaptic gaps and increased density of synaptic bodies in the ischemic hippocampal CA1 area, resulting also in increased serum GABA levels and hippocampal expressions of GABA_ARα1, SYN1 and PSD-95 and lowered expression level of CaMK II. CONCLUSIONS Electroacupuncture at Baihui and Shenting improves learning and memory function of rats with cerebral IR injury possibly through a mechanism that promotes synaptic regeneration, upregulates hippocampal expressions of GABAAR α 1, SYN1 and PSD-95 and downregulates the expression of CaMK II.
Animals ; Electroacupuncture ; Reperfusion Injury/therapy* ; Male ; Rats ; Rats, Sprague-Dawley ; Brain Ischemia/therapy* ; Hippocampus/metabolism* ; Memory ; Learning ; Disks Large Homolog 4 Protein/metabolism* ; Synapses ; Acupuncture Points ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism* ; Receptors, GABA-A/metabolism* ; gamma-Aminobutyric Acid/metabolism*

Objective: To understand and analyze the epidemiological characteristics of dental caries in children and its effects on jaw function, so as to prevent occurrence of abnormality of jaw function. Methods: In October 2020, 860 cases of children from 5 kindergartens and 3 primary schools in Beijing were selected by adopting the convenience sampling method. All of them received oral examination, and the questionnaire according to the fourth national oral health epidemiological survey standard, including demographic information, eating habits, oral hygiene habits, oral health care,etc. The epidemiological characteristics of dental caries in children and its effects on jaw function were analyzed. Results: A total of 190 children had dental caries, with a dental caries incidence rate of 22.09%. Dental caries in children s deciduous teeth or permanent teeth was related to age, residence, parents education level,whethe to eat before going to bed, whether eating sweet food, tooth brushing age, daily brushing frequency, whether parents have assisted tooth brushing, whether regular oral examination, whether parents have received oral health guidance( χ 2= 5.04 ,4.70，75.37，7.91，12.03，9.30，7.64，255.47，253.27，11.38, P <0.05). Compared with the noncaries group[(81.52±3.16， 80.54± 1.52，1.92±0.25，31.52±1.62，33.63±3.41，50.72±1.68)°], the sella nasion A point(SNA), sella nasion B point(SNB),A point nasion B point(ANB),frankfot horizontal mandibular plane angle(FH MP),skull nasion mandibular plane angle(SN MP),anteriors relationship palataplane mandibular(Ptm ANS) decreased in the dental caries group[(78.62±2.11，79.35±1.02，1.68±0.32，30.69±0.45，32.15±3.02，48.62±1.21)°](t=78.62,79.35,10.94, 30.69, 32.15, 48.62, P<0.05). Conclusion The prevalence of caries in children was high and jaw structure abnormalities and dysfunction. It is suggested that caries prevention and control work should be carried out early to strengthen the prevention and control of oral disease education work.

Long noncoding RNAs (lncRNAs) play a critical role in the regulation of atherosclerosis. Here, we investigated the role of the lncRNA growth arrest-specific 5 (lncR-GAS5) in atherogenesis. We found that the enforced expression of lncR-GAS5 contributed to the development of atherosclerosis, which presented as increased plaque size and reduced collagen content. Moreover, impaired autophagy was observed, as shown by a decreased LC3II/LC3I protein ratio and an elevated P62 level in lncR-GAS5-overexpressing human aortic endothelial cells. By contrast, lncR-GAS5 knockdown promoted autophagy. Moreover, serine/arginine-rich splicing factor 10 (SRSF10) knockdown increased the LC3II/LC3I ratio and decreased the P62 level, thus enhancing the formation of autophagic vacuoles, autolysosomes, and autophagosomes. Mechanistically, lncR-GAS5 regulated the downstream splicing factor SRSF10 to impair autophagy in the endothelium, which was reversed by the knockdown of SRSF10. Further results revealed that overexpression of the lncR-GAS5-targeted gene miR-193-5p promoted autophagy and autophagic vacuole accumulation by repressing its direct target gene, SRSF10. Notably, miR-193-5p overexpression decreased plaque size and increased collagen content. Altogether, these findings demonstrate that lncR-GAS5 partially contributes to atherogenesis and plaque instability by impairing endothelial autophagy. In conclusion, lncR-GAS5 overexpression arrested endothelial autophagy through the miR-193-5p/SRSF10 signaling pathway. Thus, miR-193-5p/SRSF10 may serve as a novel treatment target for atherosclerosis.
Humans ; Atherosclerosis/genetics* ; Autophagy/genetics* ; Cell Cycle Proteins/metabolism* ; Endothelial Cells/metabolism* ; Endothelium/metabolism* ; MicroRNAs/metabolism* ; Repressor Proteins/metabolism* ; RNA Splicing Factors ; Serine-Arginine Splicing Factors/genetics* ; RNA, Long Noncoding/metabolism*

Objective:To evaluate the effect of intraoperative individualized systolic blood pressure (SBP) management on myocardial injury after hip replacement in elderly patients at high risks of hypertension.Methods:One hundred and eighty-two patients of either sex, aged 60-89 yr, with body mass index of 18-26 kg/m 2, with a history of hypertension requiring drug treatment and stratified high risk factors of cardiovascular risk factors, scheduled for elective hip replacement under general anesthesia, were divided into 2 groups ( n=91 each) using a random number table method: routine management group and individualized SBP management group. Individualized SBP management group maintained the intraoperative SBP at 90%-110% of the baseline value, and routine management group implemented blood pressure management according to the current routine clinical pathway.The intermedian cubital venous blood samples were collected before surgery and at 24, 48, and 72 h after surgery for determination of the serum concentrations of high sensitivity cardiac troponin T. Postoperative myocardial injury and myocardial infarction were also recorded. The 30-day all-cause mortality was recorded on day 30 after surgery. Results:The incidence of postoperative myocardial injury and serum concentrations of high sensitivity cardiac troponin T at 24, 48 and 72 h after surgery were significantly decreased, and the length of hospital stay was shortened in individualized SBP management group as compared with routine management group ( P<0.05). Conclusions:Intraoperative individualized SBP management can reduce the postoperative myocardial injury in elderly patients at high risk of hypertension undergoing hip replacement.

  Objective  Different from other etiologies of early-onset scoliosis (EOS), congenital early-onset scoliosis (CEOS) is mainly linked to vertebral anomalies, such as formation failures and segmentation failures at the apex segments. So far, there is little research on CEOS patients who have completed traditional growing rods (TGR) treatment, and the initial outcomes of TGR with or without apical control technique (ACT) are different. Therefore, we compared the "final" results of CEOS patients who completed TGR treatment with or without ACT.  Methods  We conducted a retrospective study of CEOS patients who completed TGR treatment from 2007—2020. They either had final fusion or were followed up after reaching skeletal maturity. We split the patients into two groups based on whether they had ACT with TGR or not. The ACT-TGR group had apical vertebrectomy/hemivertebrectomy with short fusion and TGR. The TGR-only group had only TGR. We looked at their demographic features, radiographic measurements, and complications.  Results  This study enrolled 46 CEOS patients, of which 13 patients were in the ACT-TGR group and 33 patients in the TGR group. The ACT-TGR group had a longer distraction interval (1.17 years vs. 0.75 years). The ACT-TGR group had a larger preoperative main curve [87.00(63.50, 98.00)], but the residual curve degrees were comparable between the two groups at the last follow-up (P=0.354). At the last follow-up, the T1-12 and T1-S1 heights were similar between the two groups. The ACT-TGR group had a lower number of implant-related complications per patient (0.77 vs. 1.48). Three patients in the ACT-TGR group underwent final fusion, while 17 patients in the TGR group underwent final fusion (P=0.060).  Conclusions  Both ACT-TGR and traditional TGR coud effectively correct deformity and preserve spinal growth in CEOS patients. ACT-TGR had a better corrective effect on patients with severe deformity and did not have a significant impact on spinal height. For patients with acceptable correction, spontaneous fusion and without implant failure, retaining the implant and continuing observation could be a strategy for graduating from growing rod treatment.

Objective: To buildSERPING1-knockout porcine fetal fibroblasts(PFFs) based on CRISPR/Cas9 technology and provide cell experimental materials for the construction of hereditary angioedema models. Methods: Firstly, protein structure prediction software was used to analyze the amino acid homology between human and pigSERPING1. Secondly, the eighth exon was selected as the knockout target by screening the effective coding region of the pigSERPING1gene. A single guide RNA targeting pigSERPING1was designed, synthesized and linked to pX330 vector containing Cas9 endonuclease. G418 was used to obtain the positive monoclonal cells after transfection into PFFs. Finally, the circumstance of CRISPR/Cas9 mediated knockout was assessed by the T7 EN1 enzyme digestion assay and the genotypes of monoclonal cells were identified by sequencing analysis Results: Bioinformatic analysis revealed that theSERPING1protein of human and pig had high homology, amino acid sequence identity reached 65.87%. A vector targetingSERPING1was constructed successfully and transfected into cells. Monoclonal cells with knockoutSERPING1gene were obtained through drug screening. Sequencing confirmed the mutant genotype. Conclusion The human and pigSERPING1sequences and their protein structures are highly homologous, and it is suitable for the construction of disease model. CRISPR/Cas9 expression vectors were constructed to achieve theSERPING1gene targeting in PFFs.SERPING1-knockout monoclonal cells were obtained, which could contribute to the construction of theSERPING1knockout miniature pig model.