Objective:To explore the mechanism by which moxibustion improves cold-induced blood stasis pattern(CIBSP)and to uncover the biological basis underlying the correlation between the Conception Vessel(CV),Governor Vessel(GV),Thoroughfare Vessel(TV),and the uterus by investigating the effects of mild moxibustion on the hypothalamic-pituitary-ovary axis(HPOA)in CIBSP model rats.Methods:Forty-eight female Sprague-Dawley rats were divided into a blank group,a model group,a CV group,a GV group,a TV group,and a non-meridian non-point(NMNP)group according to the random number table method,with 8 rats in each group.Except for rats in the blank group,the other rats were subjected to an ice water bath plus epinephrine to prepare CIBSP models.Each intervention group was given mild moxibustion on the first day of modeling.Qihai(CV6),Guanyuan(CV4),and Zhongji(CV3)were selected in the CV group;Huangshu(KI16),Dahe(KI12),and Henggu(KI11)were selected in the TV group;Xuanshu(GV5),Mingmen(GV4),and Yaoyangguan(GV3)were selected in the GV group;three NMNP spots were selected in the NMNP group.Mild moxibustion was administered for 20 min each time,once a day,for 14 consecutive days.The symptom scores of CIBSP and uterine electromyogram were compared among groups;morphological changes in reproductive organs were observed by hematoxylin-eosin staining;enzyme-linked immunosorbent assay was used to detect serum reproductive hormones and neurotransmitters.The mRNA and protein expression levels of estrogen receptor(ER),follicle-stimulating hormone receptor(FSHR),and luteinizing hormone receptor(LHR)were detected by real-time quantitative polymerase chain reaction,immunohistochemistry,or Western blotting assays.Results:Compared to the blank group,the model group showed a significantly increased CIBSP symptom score,an amplified in vivo uterine myoelectric value;decreased ovarian follicles,thinner endometrial epithelium,and disordered structure with the pathological examination,decreased serum levels of reproductive hormones,6-keto-prostaglandin F1α(6-keto-PGF1α),and β-endorphin(β-EP),increased thromboxane B2(TXB2),5-hydroxytryptamine(5-HT),and substance P(SP),and decreased mRNA and protein expression of ovarian ER,FSHR,and LHR(P<0.05 or P<0.01).Compared to the model and NMNP groups,each point-intervention group showed a decreased CIBSP symptom score and uterine myoelectric value;increased ovarian follicles,thickened endometrium,increased glands,up-regulated serum reproductive hormones,6-keto-PGF1α,and β-EP,down-regulated serum TXB2,5-HT,and SP and increased mRNA and protein expression of ovarian ER,FSHR,and LHR(P<0.05 or P<0.01).Conclusion:Moxibustion at points from the CV,GV,and TV("three branches from one source")can improve the reproductive neuroendocrine function of CIBSP rats,and this effect may be mediated through the regulation of the HPOA.These detections suggest that the biological basis of point-uterus correlation may be based on HPOA regulation.

Subunit 5B of constitutively photomorphogenic 9 signalosome(CSN5B)is an inhibitory factor for the biosynthesis of vitamin C in the L-galactose synthesis pathway in plants.To create mutants with richer vitamin C in tomato fruits,a dual-target vector of pKSE402-SlCSN5Bwas constructed and trans-formed into the breeding parent of 1912.Based on the molecular biological assay and DNA sequencing,17 transgenic positive lines were determined,and 6 lines of them were genetically mutated at the target site of SlCSN5B,with an editing efficiency of 35.3％.Among the mutants,2 lines were homozygous mu-tants,csn5b-6 with 180 bp deletion and csn5b-8 with 3 bp deletion.The biological traits of two types of deficiency homozygotes without exogenous T-DNA insertion derived from the T1 generation were observed,and there were no significant differences in phenotypes of the plant and fruit.Through physiological tes-ting of red-ripe fruits from T,generation lines of csn5b-6,the GMPase activity and the vitamin C content significantly increased by 43％and 37.8％,respectively,the content of hydrogen peroxide significantly decreased by 25.9％,and the content of soluble solids did not obviously change,compared with the wild type.There were no significant differences in plant traits and physiological characteristics between T1 gen-eration lines of csn5b-8 and the wild type.Based on gene expression analysis and protein structure predic-tion,the results showed that the gene of SlCSN5B normally transcribed,and the loss of large peptide seg-ments of CSN5B encoded by SlCSN5B caused changes in its structure to affect functioning,which led to an increase of vitamin C content in the line of csn5b-6-1 1.The findings suggest that the vitamin C content of tomato fruit can be improved by CRISPR/Cas9-mediated SlCSN5B gene editing,which provides the valuable resources for high-quality breeding in tomato.

Objective To investigate the mediating effect of psychological flexibility on the relationship between pain self-efficacy and kinesiophobia in patients with lumbar disc herniation(LDH),so as to provide references for relief from kinesiophobia of the patients.Methods Convenience sampling was used to select 256 patients with LDH as the research subjects from the Outpatient Department of Spinal Orthopaedics of a Grade IIIA hospital in Guangdong Province between May and December 2023.The subjects were surveyed with a general information questionnaire,the kinesiophobia assessment scale,psychological flexibility inventory for pain patients,and chronic pain self-efficacy scale.The mediation effect of psychological flexibility on pain self-efficacy and kinesiophobia was analysed using SPSS 26.0 and the PROCESS 3.5 macro.Results The scores for kinesiophobia,psychological flexibility and pain self-efficacy among the LDH patients were 31.66±4.73,55.26±11.06 and 68.14±17.48,respectively.Kinesiophobia was positively correlated with the psychological flexibility(r=0.545,P<0.001)and negatively correlated with the pain self-efficacy(r=-0.599,P<0.001).The psychological flexibility was negatively correlated with the pain self-efficacy(r=-0.510,P<0.001).Psychological flexibility partially mediated the relationship between pain self-efficacy and kinesiophobia,with a mediating effect of-0.045,accounting for 27.78%of the total effect.Conclusion The patients who have LDH and under conservative treatment exhibit a high level of kinesiophobia and with a moderate levels of pain self-efficacy and psychological flexibility.The medical staff can improve the self-efficacy and psychological flexibility of patients,so as to reduce kinesiophobia level and its incidence.

Objective:To explore the mechanism by which moxibustion improves cold-induced blood stasis pattern(CIBSP)and to uncover the biological basis underlying the correlation between the Conception Vessel(CV),Governor Vessel(GV),Thoroughfare Vessel(TV),and the uterus by investigating the effects of mild moxibustion on the hypothalamic-pituitary-ovary axis(HPOA)in CIBSP model rats.Methods:Forty-eight female Sprague-Dawley rats were divided into a blank group,a model group,a CV group,a GV group,a TV group,and a non-meridian non-point(NMNP)group according to the random number table method,with 8 rats in each group.Except for rats in the blank group,the other rats were subjected to an ice water bath plus epinephrine to prepare CIBSP models.Each intervention group was given mild moxibustion on the first day of modeling.Qihai(CV6),Guanyuan(CV4),and Zhongji(CV3)were selected in the CV group;Huangshu(KI16),Dahe(KI12),and Henggu(KI11)were selected in the TV group;Xuanshu(GV5),Mingmen(GV4),and Yaoyangguan(GV3)were selected in the GV group;three NMNP spots were selected in the NMNP group.Mild moxibustion was administered for 20 min each time,once a day,for 14 consecutive days.The symptom scores of CIBSP and uterine electromyogram were compared among groups;morphological changes in reproductive organs were observed by hematoxylin-eosin staining;enzyme-linked immunosorbent assay was used to detect serum reproductive hormones and neurotransmitters.The mRNA and protein expression levels of estrogen receptor(ER),follicle-stimulating hormone receptor(FSHR),and luteinizing hormone receptor(LHR)were detected by real-time quantitative polymerase chain reaction,immunohistochemistry,or Western blotting assays.Results:Compared to the blank group,the model group showed a significantly increased CIBSP symptom score,an amplified in vivo uterine myoelectric value;decreased ovarian follicles,thinner endometrial epithelium,and disordered structure with the pathological examination,decreased serum levels of reproductive hormones,6-keto-prostaglandin F1α(6-keto-PGF1α),and β-endorphin(β-EP),increased thromboxane B2(TXB2),5-hydroxytryptamine(5-HT),and substance P(SP),and decreased mRNA and protein expression of ovarian ER,FSHR,and LHR(P<0.05 or P<0.01).Compared to the model and NMNP groups,each point-intervention group showed a decreased CIBSP symptom score and uterine myoelectric value;increased ovarian follicles,thickened endometrium,increased glands,up-regulated serum reproductive hormones,6-keto-PGF1α,and β-EP,down-regulated serum TXB2,5-HT,and SP and increased mRNA and protein expression of ovarian ER,FSHR,and LHR(P<0.05 or P<0.01).Conclusion:Moxibustion at points from the CV,GV,and TV("three branches from one source")can improve the reproductive neuroendocrine function of CIBSP rats,and this effect may be mediated through the regulation of the HPOA.These detections suggest that the biological basis of point-uterus correlation may be based on HPOA regulation.

Objective To investigate the mediating effect of psychological flexibility on the relationship between pain self-efficacy and kinesiophobia in patients with lumbar disc herniation(LDH),so as to provide references for relief from kinesiophobia of the patients.Methods Convenience sampling was used to select 256 patients with LDH as the research subjects from the Outpatient Department of Spinal Orthopaedics of a Grade IIIA hospital in Guangdong Province between May and December 2023.The subjects were surveyed with a general information questionnaire,the kinesiophobia assessment scale,psychological flexibility inventory for pain patients,and chronic pain self-efficacy scale.The mediation effect of psychological flexibility on pain self-efficacy and kinesiophobia was analysed using SPSS 26.0 and the PROCESS 3.5 macro.Results The scores for kinesiophobia,psychological flexibility and pain self-efficacy among the LDH patients were 31.66±4.73,55.26±11.06 and 68.14±17.48,respectively.Kinesiophobia was positively correlated with the psychological flexibility(r=0.545,P<0.001)and negatively correlated with the pain self-efficacy(r=-0.599,P<0.001).The psychological flexibility was negatively correlated with the pain self-efficacy(r=-0.510,P<0.001).Psychological flexibility partially mediated the relationship between pain self-efficacy and kinesiophobia,with a mediating effect of-0.045,accounting for 27.78%of the total effect.Conclusion The patients who have LDH and under conservative treatment exhibit a high level of kinesiophobia and with a moderate levels of pain self-efficacy and psychological flexibility.The medical staff can improve the self-efficacy and psychological flexibility of patients,so as to reduce kinesiophobia level and its incidence.

Idiopathic pulmonary fibrosis(IPF)and lung cancer(LC)pose a serious threat to human health,the prognosis of IPF is poor,the 5-year survival rate is low,and the incidence rate and mortality of LC are high.At present,IPF combined with LC(IPF-LC)has attracted much attention,but the pathogenesis is unclear.Epidemiology shows that,the risk of LC in IPF patients is 7 times than that of the general population,the incidence rate varies in different regions,which is related to smoking rate and diagnostic criteria.In terms of pathogenesis,chronic inflammation,angiogenesis,signaling pathways,aging,gene mutations,epithelial mesenchymal transition(EMT),and immune dysfunction are common pathogenic factors of both,and pathological changes such as smoking and pulmonary fibrosis increase the risk of LC.Thoroughly studying these mechanisms is of great significance for the early diagnosis,treatment strategy formulation,and improvement of patient prognosis of IPF-LC.

Pathological cardiac hypertrophy is an early and significant cardiac structural charac-teristic that contributes to the onset and progression of heart failure(HF).Its mainly structural feature is the abnormally enlarged cardiomyocyte.Effective intervention targets for abnormally en-larged cardiomyocyte remain to be identified.Previous studies have shown that the cellular shape and size can be regulated by the actin related protein 2/3(Arp2/3)complex,which is an actin-binding protein complex involved in the actin nucleation and assembly.However,the roles of the Arp2/3 complex in cardiomyocyte hypertrophy remain unknown.Here our study identifies its no-vel roles in the occurrence and development of cardiomyocyte hypertrophy.We found that mRNA levels of all subunits from the Arp2/3 complex are significantly upregulated(P<0.05)in the an-giotensin II(Ang Ⅱ)-induced neonatal rat primary and H9c2 cardiomyocyte hypertrophy.Fur-ther studies showed that siRNA-directed ARPC2 silencing inhibits the reactivation of fetal genes and enlargement of cardiomyocyte area induced by Ang Ⅱ in neonatal rat primary cardiomyocytes(NRCMs)and H9c2 cells(P<0.05).In addition,the upstream activators of the Arp2/3 com-plex including SH3 protein interacting with Nck,90 kD(SPIN90)and Ras-related C3 botulinum toxin substrate 1(Rac1)/WASp family Verprolin-homologous protein-2(WAVE-2)are upregu-lated(P<0.05)in Ang Ⅱ-induced neonatal rat primary and H9c2 cardiomyocyte hypertrophy,indicating the excessive activation of the Arp2/3 complex.We further show that CK666,a specif-ic Arp2/3 complex inhibitor,prevents the reactivation of fetal genes and the enlargement of car-diomyocyte area induced by Ang Ⅱ in NRCMs and H9c2 cells(P<0.05).Our results reveal that the Arp2/3 complex plays a crucial role in Ang Ⅱ-induced cardiomyocyte hypertrophy,which is beneficial to further studies about the molecular mechanisms by which the Arp2/3 complex regu-lates pathological cardiac hypertrophy.

Objective To observe the origin and course of the obturator artery(OA),so as to provide anatomical reference for reducing hemorrhage during pelvic surgery and pubic fracture fixation.Methods A total of 65 human hemi-pelvises specimens with intact structure were dissected to observe the origin,course and other variations of OA.Measure the length of the inner section of OA basin and the outer diameter at the origin,etc.Results OA originated from the internal iliac artery in 57 cases(87.7%),including 3 cases(4.6%)of the superior gluteal artery,5 cases(7.7%)of the inferior gluteal artery,3 cases(4.6%)of the external iliac artery and 5 cases(7.7%)of the inferior epigastric artery.OA participated in the formation of the arterial trunk in 3 cases(4.6%).The length of the pelvic segment of the OA in male and female was(50.87±15.41)mm and(51.71±14.19)mm,respectively,with no statistically significant difference between them(P>0.05).The outer diameters at the origin of the OA in male and female were(2.79±1.05)mm and(2.35±0.86)mm,and there was no statistically significant difference between them(P>0.05).Conclusion OA mainly originated from the anterior trunk of the internal iliac artery,with a few OA originated from the branches of the posterior trunk or the inferior epigastric artery,or participated in the formation of the arterial trunk.In pelvic surgery involving OA area,attention should be paid to the length of its pelvic segment and the outer diameter at the origin of OA,so as to better locate and protect blood vessels during surgery.

Aim To reveal the mechanism of CIP4 homologs protein 1(RICH1)are involved in the regu-lation of myocardial fibrosis.Methods Mouse cardiac fibroblasts(MCFs)cells were treated with transforming growth factor-β(TGF-β1)to induce the formation of a myocardial fibrosis cell model;the level of the target protein was detected by Western blotting;and the RICH1 gene was detected by transfection of the cells with plasmid.The RICH1 gene was overexpressed(RICH 1 OE)using plasmid transfection;the RICH1 gene was silenced using siRNA fragment(siRICH1);and the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3 a1,and Acta2,were de-tected using RT-qPCR.Results RICH1 was signifi-cantly down-regulated in TGF-β1-treated MCFs;the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3a1,and Acta2,were down-regu-lated in the RICH1 OE+TGF-β1 group;and in the siRICH1+TGF-β1 group,myocardial fibrosis marker genes,such as Col1 a1,Col3a1 and Acta2 were up-regulated at the expression level;phosphorylated SMAD2(p-SMAD2)and phosphorylated SMAD3(p-SMAD3)levels were down-regulated in the siRICH1 OE+TGF-β1 group.p-SMAD2 and P-SMAD3 levels were upregulated in the siRICH1+TGF-β1 group.Conclusion RICH1 inhibits TGF-β1-induced myo-cardial fibrosis;RICH1 inhibits TGF-β1-induced myo-cardial fibrosis by negatively regulating the SMAD2/3 signaling pathway.

Subunit 5B of constitutively photomorphogenic 9 signalosome(CSN5B)is an inhibitory factor for the biosynthesis of vitamin C in the L-galactose synthesis pathway in plants.To create mutants with richer vitamin C in tomato fruits,a dual-target vector of pKSE402-SlCSN5Bwas constructed and trans-formed into the breeding parent of 1912.Based on the molecular biological assay and DNA sequencing,17 transgenic positive lines were determined,and 6 lines of them were genetically mutated at the target site of SlCSN5B,with an editing efficiency of 35.3％.Among the mutants,2 lines were homozygous mu-tants,csn5b-6 with 180 bp deletion and csn5b-8 with 3 bp deletion.The biological traits of two types of deficiency homozygotes without exogenous T-DNA insertion derived from the T1 generation were observed,and there were no significant differences in phenotypes of the plant and fruit.Through physiological tes-ting of red-ripe fruits from T,generation lines of csn5b-6,the GMPase activity and the vitamin C content significantly increased by 43％and 37.8％,respectively,the content of hydrogen peroxide significantly decreased by 25.9％,and the content of soluble solids did not obviously change,compared with the wild type.There were no significant differences in plant traits and physiological characteristics between T1 gen-eration lines of csn5b-8 and the wild type.Based on gene expression analysis and protein structure predic-tion,the results showed that the gene of SlCSN5B normally transcribed,and the loss of large peptide seg-ments of CSN5B encoded by SlCSN5B caused changes in its structure to affect functioning,which led to an increase of vitamin C content in the line of csn5b-6-1 1.The findings suggest that the vitamin C content of tomato fruit can be improved by CRISPR/Cas9-mediated SlCSN5B gene editing,which provides the valuable resources for high-quality breeding in tomato.