BACKGROUND:In recent years,improving the health of skeletal muscles through exercise has become an important research concern for scholars.Appropriate exercise has a positive effect on skeletal muscles.Among them,how to activate the sphingosine kinase1(SphK1)/sphingosine-1-phase(S1P)/sphingosine-1-phase receptor2(S1PR2)signaling pathway during exercise so as to improve the health of skeletal muscles is receiving attention from researchers. OBJECTIVE:To investigate how exercise improves the health of skeletal muscles through the SphK1/S1P/S1PR2 signaling pathway,and to explore new methods for treating related muscle diseases in order to improve human skeletal muscle health. METHODS:The first author searched for relevant literature from the establishment of the database to the present in the Web of Science,PubMed,CNKI,WanFang,and VIP databases.The search terms were"signaling pathway,SphK1,S1P,S1PR2,skeletal muscle,satellite cell,myogenesis,exercise"in Chinese and English.Finally,69 articles were included for review and analysis. RESULTS AND CONCLUSION:The SphK1/S1P/S1PR2 signaling pathway is a complex regulatory network that triggers downstream signal transduction processes by SphK1 to catalyze the interaction between S1P and receptors such as S1PR2,thereby regulating multiple biological functions of cells,tissues,organs,and systems.The SphK1/S1P/S1PR2 signaling pathway can regulate satellite cell proliferation and myoblast differentiation,improving myogenesis.The physiological basis of the SphK1/S1P/S1PR2 signaling pathway and the potential impact of exercise on it were analyzed through literature research.Acute aerobic exercise can increase the expression of SphK1 in skeletal muscle.Both human and animal studies have confirmed that acute and long-term exercise can increase the expression of S1P in skeletal muscle.In addition,studies have shown that long-term resistance exercise can increase the expression of S1PR2 in skeletal muscle.Some experimental results indicate that acute and long-term exercise have no significant effect on muscle or blood S1P levels,and the reason for different results may be due to different research subjects,methods,intensities,and frequencies selected,while the specific mechanism is not yet clear.Research suggests that exercise can promote the expression of the SphK1/S1P/S1PR2 signaling pathway in skeletal muscle and regulate downstream related signaling pathways.Research on this signaling pathway may provide new strategies and methods for the treatment of skeletal muscle diseases,thereby improving skeletal muscle health.In the future,we should deepen the research on the association between SphK1/S1P/S1PR2 signaling pathway and skeletal muscle health,further reveal its regulatory relationship with satellite cells and myoblasts as well as its interactions with the upstream and downstream pathways,explore its clinical application value,take into account the changes of this pathway when formulating the rehabilitation program,explore the specific mechanisms by which different types of exercise affect the SphK1/S1P/S1PR2 signaling pathway in skeletal muscles,and use the SphK1/S1P/S1PR2 signaling pathway as a potential therapeutic target for diseases.Further development and application of human muscle models should be developed to improve research depth and accuracy.

This paper presents an automatic acquisition and analytic procedure of acupuncture manipulation based on optical navigation, aiming at solving the shortcomings of existing acquisition methods of acupuncture manipulation. An acquisition holder installed at the handle tail of filiform needle was designed to display the movement trajectory of the needle during acupuncture delivery by collecting the movement trajectory of holder. The 3-month old male Bama miniature pig was selected as the experimental subject, and 6 points, "Bojian" "Qiangfeng" "Housanli" "Xiaokua" "Huiyang" (BL35) and "Baihui" (GV20), were selected during acupuncture manipulation. The optical navigation system was used to collect the real-time data, and these data were per-processed and analyzed using mean filtering and Fourier transform. The acupuncture procedure was divided into 3 stages, inserting, lifting-thrusting, and twisting. The results showed that the accuracy was 96.3% at lifting-thrusting stage, and that was 100.0% at twisting stage. The decomposition effect of the entire procedure was satisfactory. This study provides a new approach to the quantitative analysis of acupuncture manipulation. In the future, it needs to further optimize the algorithm and expand the sample size so as to improve the accuracy of this analytic technique.
Acupuncture Therapy/methods* ; Male ; Animals ; Swine ; Acupuncture Points ; Humans ; Swine, Miniature ; Needles

This study explored whether Astragalus polysaccharide(APS)can regulate the VEGF/VEGFR signaling pathway to affect the proliferative activity of rat intestinal mucosal microvascu-lar endothelial cells(RIMMVECs).RIMMVECs were isolated from newborn rats,then purified and treated with APS at concentrations of 0.1,1.0,10.0,100.0,1 000.0,and 10 000.0 mg/L.MTT was used to determine the effect of APS on RIMMVECs proliferation and screen for the optimal concentration of APS.Subsequently,flow cytometry was used to detect the changes in cell cycle to evaluate the stage of action of APS on the cell cycle in RIMMVECs.Then,the ELISA was used to detect the changes of VEGFA in cell supernatant to evaluate the potential of cell proliferation and angiogenesis.The changes in fluorescence intensity of Fluo-8AM was observed using fluorescence microscopy to evaluate intracellular Ca2+levels.Finally,Western blot was used to detect the ex-pression of PERK in RIMMVECs to analyze the possible mechanism of APS.The results showed that 100 mg/L APS significantly enhanced the proliferative activity of RIMMVECs,increased the content of VEGFA in the cell supernatant,the intracellular Ca2+levels,and the expression of PERK protein,indicating that APS promotes the proliferation of RIMMVECs,which may be a-chieved by promoting the expression of VEGFA and activating the ERK pathway.

This study explored whether Astragalus polysaccharide(APS)can regulate the VEGF/VEGFR signaling pathway to affect the proliferative activity of rat intestinal mucosal microvascu-lar endothelial cells(RIMMVECs).RIMMVECs were isolated from newborn rats,then purified and treated with APS at concentrations of 0.1,1.0,10.0,100.0,1 000.0,and 10 000.0 mg/L.MTT was used to determine the effect of APS on RIMMVECs proliferation and screen for the optimal concentration of APS.Subsequently,flow cytometry was used to detect the changes in cell cycle to evaluate the stage of action of APS on the cell cycle in RIMMVECs.Then,the ELISA was used to detect the changes of VEGFA in cell supernatant to evaluate the potential of cell proliferation and angiogenesis.The changes in fluorescence intensity of Fluo-8AM was observed using fluorescence microscopy to evaluate intracellular Ca2+levels.Finally,Western blot was used to detect the ex-pression of PERK in RIMMVECs to analyze the possible mechanism of APS.The results showed that 100 mg/L APS significantly enhanced the proliferative activity of RIMMVECs,increased the content of VEGFA in the cell supernatant,the intracellular Ca2+levels,and the expression of PERK protein,indicating that APS promotes the proliferation of RIMMVECs,which may be a-chieved by promoting the expression of VEGFA and activating the ERK pathway.

Objective To investigate the role of fragile X mental retardation protein(FMRP)in promoting cell migration and epithelial-mesenchymal transition(EMT)in breast cancer(BC)and the potential mechanisms involved.Methods The mRNA and protein expressions of FMRP in MCF-10A,a normal human breast epithelial cell line,and four breast cancer cell lines,including MCF-7,BT474,MDA-MB-231,and HCC1937,were analyzed by RT-PCR and Western blot.The expression of FMRP in BC tissues was measured by immunohistochemistry(IHC).FMRP expression in BC and its relationship with clinical prognosis were analyzed using GEO database.Lentiviral infection and siRNA interference were used to construct FMRP overexpression and interference vectors,respectively,and the human breast cancer cell line MCF-7 was subsequently transfected.A Control group,an interference empty vector group(the NC group),a knockdown vector group(the siFMRP group),an overexpression empty vector group(the Lv-NC group),and an overexpression vector group(the Lv-FMRP group)were set up.The migration and invasion abilities of the cells were assessed by scratch assay and Transwell assay.The expression of EMT markers,including E-cadherin,an epithelial marker,N-cadherin,an mesenchymal markers,vimentin,zinc finger E-box binding homeobox 1(ZEB1),and snail family zinc finger 2(Slug),in the cells of each group was determined by Western blot.The interaction between FMRP and DEAD-box RNA helicase-5(DDX5)protein was analyzed by immunocoprecipitation combined with mass spectrometry(IP-MS).The regulatory effect of FMRP on DDX5 protein expression was assessed using the protein synthesis inhibitor cycloheximide(CHX)and proteasome inhibitor MG132.In addition,transfection with siDDX5 vector was conducted to observe whether DDX5 could reverse the effects of FMRP overexpression on cell migration and EMT.The localization and expression of β-catenin were determined by immunofluorescence staining,and the expression of core markers of Wnt/β-catenin signaling pathway was examined by Western blot.Results FMRP was highly expressed in BC tissues and cells(P＜0.05),and overall survival(OS)and recurrence-free survival(RFS)of the FMRP high expression group were significantly lower than those of the FMRP low expression group(P＜0.05).The migration ability of MCF-7 cells was weakened after FMRP knockdown,while overexpression of FMRP promoted cell migration(P＜0.05).After FMRP knockdown,the expression of E-cadherin was increased,while the expression levels of N-cadherin,vimentin,ZEB1,and Slug were decreased,which inhibited the occurrence of EMT.In contrast,the overexpression of FMRP promoted the EMT process(P＜0.05).FMRP interacted with DDX5 protein and promoted DDX5 protein stability by blocking the ubiquitin-proteasome pathway.DDX5 knockdown reversed the effect of FMRP overexpression to promote cell migration and EMT(P＜0.05),effectively inhibited β-catenin nuclear translocation,and decreased β-catenin nuclear distribution.Furthermore,it was found that the expression of p-β-catenin,GSK3β and Axin2 protein was increased and the expression of C-myc protein was decreased after DDX5 downregulation(P＜0.05).On the other hand,the expression of these proteins was reversed by combined FMRP overexpression(P＜0.05).Conclusion FMRP targets DDX5 and promotes BC cell migration and EMT via the activation of the Wnt/β-catenin signaling pathway.

Objective:To investigate the surgical technique and clinical effect of the island flap of dorsal branch of proper palmar digital artery in repair of the soft tissue defect of digits.Methods:From March 2013 to March 2021, 22 cases of digit soft tissue defects were repaired with dorsal branch of proper palmar digital artery island flap. The digit defects involved: 9 thumbs, 5 index fingers, 3 middle fingers, 3 ring fingers and 2 little fingers. The repair of defects covered 8 digit-tips, 7 pulps and 7 dorsal and nail beds. The defected area of soft tissue was 0.8 cm×0.5 cm-1.5 cm×8.0 cm, and the size of flap was 1.0 cm×0.7 cm-1.8 cm×1.0 cm. The donor site in 6 cases was closed directly. The other 16 cases were covered with medium thickness skin graft and pressurised bandaging. The follow-up reviews were carried out via the outpatient clinic visit, telephone or WeChat interview. Results:After operation, 1 flap had cyanosis due to a tight suture and it was relieved after the removal of intermittent suture; Tension blisters appeared in 2 cases and disappeared after 1 week; One case had necrosis at distal flap and healed after dressing change. Other flaps survived successfully and the incision and donor site healed in the first stage. All patients were entered to 6 to 18(mean 10) months of follow-up. At the final follow-up, the appearance and texture of the flaps were good and the protective sensation was restored. The flexion and extension function of the affected digit was normal with the TPD at 7-11 mm. The original shape and function of the digit body were basically reconstructed, except the failure in reconstruction of the special structure of digit body, such as nail, finger pulp thread and fine sensation. According to the Evaluation Standard of Upper Limb Function of Chinese Hand Surgery Society, 11 cases were in excellent, 9 in good and 2 in fair. The excellent and good rate was 91%. The function at donor sites was not affected.Conclusion:Repair of digit soft tissue defect with dorsal branch of proper palmar digital artery island flap is easy to operate, and with a low risk, high success rate and satisfactory curative effect.

Objective:To investigate the dosimetric differences between conventional IMRT and electron beam conformal radiotherapy (EBCRT) combined with IMRT for post-mastectomy left-sided breast cancer patients.Methods:A total of 20 post-mastectomy left-sided breast cancer patients who were treated in the Ningbo First Hospital from June 2018 to October 2021 were retrospectively studied. The planning target volume (PTV) included the supra-and infra-clavicular regions(PTV sc)and the ipsilateral chest wall (PTV cw), and the prescribed dose was 50 Gy/25 f. All radiotherapy plans were designed using the Varian Eclipse treatment planning system (TPS). After that, the dose distribution of the target volume and the dose exposure of organs at risk (OARs) were compared and analyzed. Results:All the IMRT plans met the clinical requirements, yet 2/20 of the EBCRT combined with IMRT plans were not clinically accepted. For these two patients, the maximum chest wall thickness was 3.7 cm and 4.4 cm each, and the designed electron beam energy was 12 MeV and 15 MeV, respectively. The dose to the ipsilateral lung of these two patients exceeded the institution-specific dose limit standard. For the remaining 18 patients whose chest wall thickness was 3 cm or less, the designed electron beams were 9 MeV or less. All the EBCRT combined with IMRT plans were clinically accepted. The target dose distribution of the conventional IMRT was better than that of the EBCRT combined with IMRT (uniformity index (HI): PTV sc: t = -10.20, P<0.05; PTV cw: t = -9.24, P<0.05; conformal index (CI): PTV all: t = 10.39, P <0.05). For OARs, the V5 Gy, V20 Gy, and Dmean of the ipsilateral lung of EBCRT combined with IMRT were lower than those of IMRT ( t = 5.98, 6.30, 11.30, P <0.05). Specifically, the V25 Gy and Dmean of heart decreased by 8.3% and 4.79 Gy, respectively ( t = 15.23, 15.76, P<0.05), the Dmean of the left anterior descending coronary artery (LADCA) decreased by 44.03% ( t = 11.69, P <0.05), and the V5 Gy and Dmean of the contralateral breast decreased by 7.9% and 0.8 Gy, respectively ( t = 3.66, 4.93, P<0.05). The dosimetric differences of other OARs were not statistically significant ( P > 0.05). Conclusions:For post-mastectomy left-sided breast cancer patients with a chest wall thickness of less than 3 cm, EBCRT combined IMRT can significantly reduce the exposure dose to the heart, the ipsilateral lung, and the contralateral breast, which is beneficial to reducing the potential risk of long-term complications after radiotherapy and can further improve the long-term overall survival rate of patients. For patients with thick chest wall, IMRT plans are more technologically ideal.

Objective: To detect potential mutation in a Chinese pedigree affected with congenital limb malformations. Methods: Clinical data was collected. Genomic DNA was extracted from peripheral blood samples of family members. The zone of polarizing activity regulatory sequence (ZRS) were amplified by PCR and subjected to direct sequencing. Results: Among the 13 individuals in this pedigree, there were 4 PPD patients, who were characterized by varying degrees of deformity. The female patients suffered triphalangeal thumb and preaxial polydactyly, while the male patients only had preaxial polydactyly. Only one patient had foot involvement. TA heterogeneous mutations was discovered in the ZRS (105C>G) in all patients, the same mutation was not detected in 2 healthy family members. Conclusion The inheritance pattern of PPD was autosomal dominant inheritance. There was a significant variability of symptoms among family patients. The heterozygous mutation of the ZRS (105C>G) probably underlie the disease.

OBJECTIVE: To detect potential mutation in a Chinese pedigree affected with congenital limb malformations. METHODS: Clinical data was collected. Genomic DNA was extracted from peripheral blood samples of family members. The zone of polarizing activity regulatory sequence (ZRS) were amplified by PCR and subjected to direct sequencing. RESULTS: Among the 13 individuals in this pedigree, there were 4 PPD patients, who were characterized by varying degrees of deformity. The female patients suffered triphalangeal thumb and preaxial polydactyly, while the male patients only had preaxial polydactyly. Only one patient had foot involvement. TA heterogeneous mutations was discovered in the ZRS (105C>G) in all patients, the same mutation was not detected in 2 healthy family members. CONCLUSION The inheritance pattern of PPD was autosomal dominant inheritance. There was a significant variability of symptoms among family patients. The heterozygous mutation of the ZRS (105C>G) probably underlie the disease.
Female ; Genetic Testing ; Hand Deformities, Congenital ; genetics ; Humans ; Limb Deformities, Congenital ; genetics ; Male ; Membrane Proteins ; genetics ; Pedigree ; Polydactyly ; genetics ; Thumb ; pathology

Objective To study the effect of multi-point multi-axis suspension training on nonspecific low back pain (NLBP) through surface electromyography (sEMG). Methods From October, 2016 to November, 2017, 24 patients with unilateral NLBP (NLBP group) were randomly divided into dynamic group (n=12) and static group (n=12); other 12 healthy volunteers were selected as control group. NLBP group was trained on multi-point multi-axis suspension training system, the dynamic group accepted pelvic-up training in the supine position, while the static group accepted lumbar neutral keep training, for ten days. NLBP group was measured with average electromyographic values (AEMG) of affected musculi multifidi before and after treatment, and assessed with Visual Analogue Scale (VAS) of pain; while the control group was measured AEMG of unilateral musculi multifidi. Results The AEMG in static and pelvic-up positions was more in NLBP group than in the control group before treatment (t>3.209, P<0.01), and it was less when keeping neutral position (t=-2.364, P<0.05). The AEMG improved in NLBP groups after treatment (t>2.982, P<0.01). The AEMG in static position and in keeping neutral position was not significantly different among the three groups after treatment (F<2.921, P>0.05), and it was the most in the dynamic group as pelvic-up (P<0.001). The score of VAS decreased in both NLBP groups after treatment (t>10.416, P<0.001), and it was less in the static group than in the dynamic group (t=-4.389, P<0.001).Conclusion sEMG changes take place in patients with NLBP. Multi-point multi-axis suspension training is effective through improving the core muscle group, and it is more effective in static maintenance training.