Objective: To investigate the effects of “Three Methods and Three Acupoints” (TMTP) Tuina therapy on spinal microcirculation in sciatic nerve injury (SNI). Methods: Thirty-six Sprague–Dawley rats were randomly assigned to four groups: normal, sham operation, model, and TMTP Tuina. Successful model induction was confirmed by observable hind limb lameness. After 20 sessions, hind limb grip strength and motor nerve conduction velocity (MNCV) were measured at baseline and following the 10th and 20th intervention. CD31 and α-SMA in the ventral horn of SNI model rats were detected using immunofluorescence. Motor neurons in the ventral horn were detected by Nissl staining. PTEN levels in the ventral horn were measured by ELISA, and PI3K, Akt, BDNF, VEGF, and HIF-1α expression was determined by RT-PCR. Spinal cord microcirculation was evaluated by western blotting analysis of the levels of Akt, p-Akt, BDNF, and VEGF. Results: Hind limb grip strength and MNCV significantly improved in the TMTP Tuina group compared to the model group (both P < .001). Morphology of ventral horn motor neurons in the TMTP Tuina group improved compared to the model group, with increased expressions of α-SMA (P = .002) and CD31 (P = .006). Western blot analysis indicated increased expression of VEGF (P = .005), p-Akt (P < .001), and BDNF (P = .008) in the ventral horn following Tuina treatment. RT-PCR analysis revealed increased expression of PI3K, Akt, BDNF, VEGF and HIF-1α (all P < .05). In contrast, expression of PTEN decreased compared to the model group (P < .001). Conclusion TMTP Tuina therapy may restore motor function in rats, enhance ventral horn motor neuron morphology, and promote angiogenesis and vascular smooth muscle proliferation. The mechanism may involve the activation of the PI3K/Akt signaling pathway.

Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors,including non-small cell lung cancer(NSCLC).However,its detailed molecular mechanism has not been adequately demonstrated.In this research,it was demonstrated that elemene effectively curtailed NSCLC growth in the patient-derived xenograft(PDX)model.Mechanistically,employing high-throughput screening techniques and subsequent biochemical validations such as microscale thermophoresis(MST),microRNA-145-5p(miR-145-5p)was pinpointed as a critical target through which elemene exerts its anti-tumor effects.Inter-estingly,elemene serves as a binding stabilizer for miR-145-5p,demonstrating a strong binding affinity(dissociation constant(KD)=0.39±0.17 μg/mL)and preventing its degradation both in vitro and in vivo,while not interfering with the synthesis of the primary microRNA transcripts(pri-miRNAs)and precursor miRNAs(pre-miRNAs).The stabilization of miR-145-5p by elemene resulted in an increased level of this miRNA,subsequently suppressing NSCLC progression through the miR-145-5p/mitogen-activated pro-tein kinase kinase kinase 3(MAP3K3)/nuclear factor kappaB(NF-κB)pathway.Our findings provide a new perspective on revealing the interaction patterns between clinical anti-tumor drugs and miRNAs.

Elemene is widely recognized as an effective anti-cancer compound and is routinely administered in Chinese clinical settings for the management of several solid tumors, including non-small cell lung cancer (NSCLC). However, its detailed molecular mechanism has not been adequately demonstrated. In this research, it was demonstrated that elemene effectively curtailed NSCLC growth in the patient-derived xenograft (PDX) model. Mechanistically, employing high-throughput screening techniques and subsequent biochemical validations such as microscale thermophoresis (MST), microRNA-145-5p (miR-145-5p) was pinpointed as a critical target through which elemene exerts its anti-tumor effects. Interestingly, elemene serves as a binding stabilizer for miR-145-5p, demonstrating a strong binding affinity (dissociation constant (K _D) = 0.39 ± 0.17 μg/mL) and preventing its degradation both in vitro and in vivo, while not interfering with the synthesis of the primary microRNA transcripts (pri-miRNAs) and precursor miRNAs (pre-miRNAs). The stabilization of miR-145-5p by elemene resulted in an increased level of this miRNA, subsequently suppressing NSCLC progression through the miR-145-5p/mitogen-activated protein kinase kinase kinase 3 (MAP3K3)/nuclear factor kappaB (NF-κB) pathway. Our findings provide a new perspective on revealing the interaction patterns between clinical anti-tumor drugs and miRNAs.

Objective To observe the effects of tuina of"three methods and three acupoints"on skeletal muscle α-actin,myostatin(MSTN)and atrophy gene 1(Atrogin1)expression of sciatic nerve injury(SNI)rats;To explore the mechanism of tuina therapy on motor dysfunction.Methods Male SD rats were randomly divided into blank group,sham-operation group,model group and tuina group,with 9 rats in each group.SNI model was established by clamp method in rats of the model group and tuina group.The sciatic nerve was exposed without clamping in rats of the sham-operation group,the blank group was not intervened.7 days after the operation,the intelligent tuina manipulation simulator was used to simulate the point method,dial method and knead method,which were applied to the"Yinmen"(BL37),"Chengshan"(BL57)and"Yanglingquan"(GB34)of rats in the tuina group,once a day,for 20 times.The rats in the sham-operation group and the model group were only grasped and restrained.Rats in the blank group did not receive any intervention.The hind limb muscle strength were evaluated by inclined plate test before modeling,after 10 interventions and 20 interventions.After the intervention,the rats were euthanized.The expressions of α-actin in gastrocnemius muscle tissue were detected by immunofluorescence staining,the expressions of MSTN,Atrogin1 mRNA and protein in gastrocnemius muscle tissue were detected by RT-PCR and Western blot.Results Compared with the blank group and sham-operation group,the model group showed a decrease in hind limb muscle strength(P<0.01),a significant decrease in α-actin expression in gastrocnemius muscle tissue(P<0.01),and a significant increase in MSTN,Atrogen1 mRNA and protein expression(P<0.05,P<0.01).Compared with the model group,the hind limb muscle strength in tuina group significantly increased(P<0.01),the expressions of α-actin significantly increased(P<0.01),and the expressions of MSTN,Atrogin1 mRNA and protein significantly decreased(P<0.05,P<0.01).Conclusion"Three methods and three acupoints"tuina can improve hind limb muscle strength and restore motor function of SNI rats,which is related to the down-regulation of MSTN and Atrogin1 as well as increasing the expression of α-actin in gastrocnemius muscles.

AIM: To explore the risk factors for visual field defect in patients with normal tension glaucoma.METHODS: The general data of 164 patients(164 eyes)with normal tension glaucoma diagnosed and treated in our hospital from July 2022 to January 2025 were retrospectively analyzed. According to the visual field defect, the patients were divided into visual field defect group(93 eyes)and no visual field defect group(71 eyes). The clinical data of the two groups were compared, and the risk factors of visual field defect in patients with normal intraocular pressure glaucoma were analyzed by Logistic regression.RESULTS: Totally 93 eyes had visual field defect among 164 eyes with normal tension glaucoma, with an incidence rate of 57%. The age, proportion of high myopia, intraocular pressure, intraocular pressure fluctuation and resistance index(RI)in the visual field defect group were higher than those in the non-visual field defect group(all P<0.01), while the retinal nerve fiber layer thickness, the optic disc vascular density, peak systolic blood flow velocity(PSV)and end diastolic blood flow velocity(EDV)in the visual field defect group were lower than those in the non-visual field defect group(all P<0.01). Multivariate Logistic regression showed that advanced age(OR=1.171), high myopia(OR=2.316), ocular hypertension(OR=1.662), high intraocular pressure fluctuation(OR=1.770), low retinal nerve fiber layer thickness(OR=0.744), low optic disc vascular density(OR=0.547), low PSV(OR=0.618), low EDV(OR=0.577)and high RI(OR=1.980)were all risk factors for visual field defect in patients with normal tension glaucoma(all P<0.01).CONCLUSION: The age, high myopia, intraocular pressure, intraocular pressure fluctuation, retinal nerve fiber layer thickness, optic disc vascular density, PSV, EDV and RI are influence factors for visual field defect in patients with normal tension glaucoma.

The cellular and molecular components of the tumor immune microenvironment (TIME) and their information exchange processes significantly influence the trends of anti-tumor immunity. In recent years, numerous studies have begun to evaluate TIME in the context of previous cancer treatment strategies. This review will systematically summarize the compositional characteristics of TIME and, based on this foundation, explore the impact of current cancer therapies on the remodeling of TIME, aiming to provide new insights for the development of innovative immune combination therapies that can convert TIME into an anti-tumor profile.
Tumor Microenvironment/immunology* ; Humans ; Neoplasms/therapy* ; Immunotherapy/methods* ; Animals

Objective To observe the effects of tuina of"three methods and three acupoints"on skeletal muscle α-actin,myostatin(MSTN)and atrophy gene 1(Atrogin1)expression of sciatic nerve injury(SNI)rats;To explore the mechanism of tuina therapy on motor dysfunction.Methods Male SD rats were randomly divided into blank group,sham-operation group,model group and tuina group,with 9 rats in each group.SNI model was established by clamp method in rats of the model group and tuina group.The sciatic nerve was exposed without clamping in rats of the sham-operation group,the blank group was not intervened.7 days after the operation,the intelligent tuina manipulation simulator was used to simulate the point method,dial method and knead method,which were applied to the"Yinmen"(BL37),"Chengshan"(BL57)and"Yanglingquan"(GB34)of rats in the tuina group,once a day,for 20 times.The rats in the sham-operation group and the model group were only grasped and restrained.Rats in the blank group did not receive any intervention.The hind limb muscle strength were evaluated by inclined plate test before modeling,after 10 interventions and 20 interventions.After the intervention,the rats were euthanized.The expressions of α-actin in gastrocnemius muscle tissue were detected by immunofluorescence staining,the expressions of MSTN,Atrogin1 mRNA and protein in gastrocnemius muscle tissue were detected by RT-PCR and Western blot.Results Compared with the blank group and sham-operation group,the model group showed a decrease in hind limb muscle strength(P<0.01),a significant decrease in α-actin expression in gastrocnemius muscle tissue(P<0.01),and a significant increase in MSTN,Atrogen1 mRNA and protein expression(P<0.05,P<0.01).Compared with the model group,the hind limb muscle strength in tuina group significantly increased(P<0.01),the expressions of α-actin significantly increased(P<0.01),and the expressions of MSTN,Atrogin1 mRNA and protein significantly decreased(P<0.05,P<0.01).Conclusion"Three methods and three acupoints"tuina can improve hind limb muscle strength and restore motor function of SNI rats,which is related to the down-regulation of MSTN and Atrogin1 as well as increasing the expression of α-actin in gastrocnemius muscles.

Objective By observing the effects of"three methods and three points"tuina technique on the expression of interleukin-17F(IL-17F),interleukin-17 receptor C(IL-17RC),activator 1 of nuclear transcription factor-κB(Act1),tumour necrosis factor receptor-associated factor 6(TRAF6)and M1 microglial cell expression in the spinal dorsal horn of rats with mild chronic compressive injury(minor CCI)model,we explored the immediate analgesic mechanism of tuina on peripheral neuropathic pain(pNP).Methods Thirty-six SD rats were divided into the sham group,the model group and the tuina group according to the random number method,twelve rats in each group,and the minor CCI model was replicated by ligating the right sciatic nerve.The rats in the tuina group were subjected to pointing,plucking and kneading at the BL37,BL57 and GB34 points on the affected side using a tuina simulator,while the sham group and the model group were only grasped and restrained,and were intervened for one time.The mechanical pain test and cold plate test were used to evaluate the response of rats to mechanical stimulation and cold stimulation after immediate intervention.The protein expression of IL-17F and TRAF6 in the spinal dorsal horn of rats in each group was detected by Western blotting.The mRNA expression of IL-17F,IL-17RC,Act1 and TRAF6 in the spinal dorsal horn of rats in each group was detected by real-time PCR.The average fluorescence intensity of M1 microglia in the spinal dorsal horn of rats in each group was detected by immunofluorescence.Results Behavioral results showed that before intervention,compared with the sham group,paw mechanical withdraw threshold(PMWT)decreased and cold sensitivity threshold(CST)increased in the model group and the tuina group;after tuina intervention,PMWT in the tuina group was increased,and CST was decreased compared with the model group;after intervention,PMWT in the tuina group was increased,while CST was decreased(P＜0.05).RT-PCR results showed that compared with the sham group,mRNA expression levels of IL-17F,IL-17RC,TRAF6 and Act1 in the spinal dorsal horn of the model group were increased;compared with model group,the mRNA expression levels of above indexes in the tuina group were decreased(P＜0.05).Western boltting results showed that compared with the sham group,the expression levels of IL-17F and TRAF6 protein in the spinal dorsal horn of the model group were increased;compared with the model group,the expression levels of IL-17F and TRAF6 protein in the tuina group decreased(P＜O.05).Immunofluorescence results showed that the mean fluorescence intensity of CD40 in the spinal dorsal horn of model group was enhanced compared with the sham group;compared with the model group,the mean fluorescence intensity of CD40 in the tuina group was decreased(P＜0.05).Conclusion The"three methods and three points"tuina technique can produce immediate analgesia by inhibiting the expression of IL-17F,IL-17RC,Act1,TRAF6 and the activation of M1 microglia in the dorsal horn of the spinal cord after one intervention.

Objective To explore the analgesic initiation mechanism of"three-manipulations and three-acupoints"of tuina on minor chronic constriction injury(minor CCI)model rats.Methods According to the random number table method,35 SD rats were randomly divided into five groups:normal group,sham group,model group,tuina group,and tuina+MK-801 group.The model group,tuina group,and tuina+MK-801 group were subjected to ligation of the right sciatic nerve trunk to establish a minor CCI rat model.The sham group was only exposed to the right sciatic nerve without ligation,and the normal group was not subjected to any operation.The normal group was not subjected to any intervention measures.On the seventh day after modeling,the model group and the sham group underwent 9 minutes of grasping restraint,while the tuina group underwent one intervention of three-manipulations(point method,dialing method,and kneading method)and three-acupoints(right"Yinmen"(BL37),"Chengshan"(BL57),and"Yanglingquan"(GB34)acupoints)with each manipulation and acupoint intervention for 1 minute for a total of 9 minutes.The tuina+MK-801 group received intrathecal injection of MK-801 from the fifth to seventh days after modeling,with a dose of 6 μg(10 μL)per day,tuina intervention was performed 30 minutes after the last intrathecal injection,and the specific operation of tuina was the same as that of the tuina group.Before modeling,after modeling,and after intervention,each group of rats was subjected to cold sensitivity threshold(CST)and mechanical withdrawal threshold(MWT)testing.After intervention,immunohistochemistry was used to detect the positive expression of cyclic guanosine monophosphate(cGMP)in the spinal dorsal horn(SDH)at L4-6 segments;protein expressions of N-methyl-D-aspartate receptor 1(NMDAR1),neurogenic nitric oxide synthase(nNOS),soluble guanylyl cyclase β(sGCβ),and protein kinase G1(PKG1)in SDH at L4-6 segments were detected by Western blotting;mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 in SDH at L4-6 segments were detected by real-time PCR.Results Compared with the normal and sham groups,after modeling,CST increased and MWT decreased in the model group,tuina group and tuina+MK-801 group(P＜0.05);after intervention,the positive protein expression of cGMP was increased,the protein expressions of NMDAR1,nNOS,sGCβ,and PKG1 were increased,and mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 were increased in SDH at L4-6 segments in the model group(P＜0.05).Compared with the model group,after intervention,CST decreased and MWT increased in the tuina group and tuina+MK-801 group(P＜0.05);the positive protein expression of cGMP was decreased,the protein expressions of NMDAR1,nNOS,sGCβ,and PKG1 were decreased,and mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 were decreased in SDH at L4-6 segments in the tuina group and tuina+MK-801 group(P＜0.05).Conclusion One-time tuina intervention can effectively improve the symptoms of thermal and mechanical hyperalgesia induced by peripheral nerve injury,which may initiate analgesia through the NMDAR1/cGMP/protein kinase G signaling pathway,thereby exerting immediate analgesic effect.

Background::The conversion of adenosine (A) to inosine (I) through deamination is the prevailing form of RNA editing, impacting numerous nuclear and cytoplasmic transcripts across various eukaryotic species. Millions of high-confidence RNA editing sites have been identified and integrated into various RNA databases, providing a convenient platform for the rapid identification of key drivers of cancer and potential therapeutic targets. However, the available database for integration of RNA editing in hematopoietic cells and hematopoietic malignancies is still lacking.Methods::We downloaded RNA sequencing (RNA-seq) data of 29 leukemia patients and 19 healthy donors from National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database, and RNA-seq data of 12 mouse hematopoietic cell populations obtained from our previous research were also used. We performed sequence alignment, identified RNA editing sites, and obtained characteristic editing sites related to normal hematopoietic development and abnormal editing sites associated with hematologic diseases.Results::We established a new database, "REDH", represents RNA editome in hematopoietic differentiation and malignancy. REDH is a curated database of associations between RNA editome and hematopoiesis. REDH integrates 30,796 editing sites from 12 murine adult hematopoietic cell populations and systematically characterizes more than 400,000 edited events in malignant hematopoietic samples from 48 cohorts (human). Through the Differentiation, Disease, Enrichment, and knowledge modules, each A-to-I editing site is systematically integrated, including its distribution throughout the genome, its clinical information (human sample), and functional editing sites under physiological and pathological conditions. Furthermore, REDH compares the similarities and differences of editing sites between different hematologic malignancies and healthy control.Conclusions::REDH is accessible at http://www.redhdatabase.com/. This user-friendly database would aid in understanding the mechanisms of RNA editing in hematopoietic differentiation and malignancies. It provides a set of data related to the maintenance of hematopoietic homeostasis and identifying potential therapeutic targets in malignancies.