BACKGROUND:Carnosic acid,a bioactive compound found in rosemary,has been shown to reduce inflammation and reactive oxygen species(ROS).However,its mechanism of action in osteoclast differentiation remains unclear. OBJECTIVE:To investigate the effects of carnosic acid on osteoclast activation,ROS production,and mitochondrial function. METHODS:Primary bone marrow-derived macrophages from mice were extracted and cultured in vitro.Different concentrations of carnosic acid(0,10,15,20,25 and 30 μmol/L)were tested for their effects on bone marrow-derived macrophage proliferation and toxicity using the cell counting kit-8 cell viability assay to determine a safe concentration.Bone marrow-derived macrophages were cultured in graded concentrations and induced by receptor activator of nuclear factor-κB ligand for osteoclast differentiation for 5-7 days.The effects of carnosic acid on osteoclast differentiation and function were then observed through tartrate-resistant acid phosphatase staining,F-actin staining,H2DCFDA probe and mitochondrial ROS,and Mito-Tracker fluorescence detection.Western blot and RT-PCR assays were subsequently conducted to examine the effects of carnosic acid on the upstream and downstream proteins of the receptor activator of nuclear factor-κB ligand-induced MAPK signaling pathway. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and F-actin staining showed that carnosic acid dose-dependently inhibited in vitro osteoclast differentiation and actin ring formation in the cell cytoskeleton,with the highest inhibitory effect observed in the high concentration group(30 μmol/L).Carnosic acid exhibited the most significant inhibitory effect during the early stages(days 1-3)of osteoclast differentiation compared to other intervention periods.Fluorescence imaging using the H2DCFDA probe,mitochondrial ROS,and Mito-Tracker demonstrated that carnosic acid inhibited cellular and mitochondrial ROS production while reducing mitochondrial membrane potential,thereby influencing mitochondrial function.The results of western blot and RT-PCR revealed that carnosic acid could suppress the expression of NFATc1,CTSK,MMP9,and C-fos proteins associated with osteoclast differentiation,and downregulate the expression of NFATc1,Atp6vod2,ACP5,CTSK,and C-fos genes related to osteoclast differentiation.Furthermore,carnosic acid enhanced the expression of antioxidant enzyme proteins and reduced the generation of ROS during the process of osteoclast differentiation.Overall,carnosic acid exerts its inhibitory effects on osteoclast differentiation by inhibiting the phosphorylation modification of the P38/ERK/JNK protein and activating the MAPK signaling pathway in bone marrow-derived macrophages.

Objective To explore the differences in stress distribution and stability of the planta pedis in the patients with unilateral and bilateral plantar fasciitis(PF)through plantar stress and center of pressure(COP)analysis.Methods A total of 100 patients with PF visiting in this hospital were enrolled,among them 50 cases were unilateral heel pain(unilateral heel pain group)and 50 cases were bilateral heel pain(bilateral heel pain).Meanwhile,50 healthy subjects were included(healthy group).In the health group and bilateral heel pain groups,the average stress value of both planta pedis surfaces of each subject was taken and named as the J0 group and H2 group,respectively.In the unilateral heel pain group,the plantar stress in 50 healthy feet and 50 affected feet were named as the J1 group and H1 group,respectively.The plantar pedis was divided into 10 regions for analysis and comparison[the first foot toe(T1),T2-5,the first-fifth metatarsal bones(M1-M5),the mid foot(MF),heel medial side(MH),heel lateral side(LH)].The subjects in 3 groups conducted the static and dynamic tests respectively,and the differences in plantar stress distribution and COP parameters among the J1,H1,H2 and J0 groups were compared respectively.Results In the static tests,the maximum pressure of the LH regions in the group J1 was increased when compared with the group J0,the contact area of LH regions in the group H1 was reduced when compared with the group J0,the maximum pressure of the M2 and M3 regions in the H1 group was increased when compared with the group J0,the contact areas of the MH and LH regions in the H2 group were decreased when compared with the group J0,the maximum pressure of the M1 region was increased when compared with the group J0,and the differences were statistically significant(P<0.05).In the dynamic tests,the maximum pressure of the T2-5 regions in the J1 group was increased when compared with the J0 group,the maximum pressure of the M3 region in the group H1 was increased when compared with the group J0,the maximum pressure of the M3 and M4 region in the group H2 was in-creased when compared with the group J0,and the differences were statistically significant(P<0.05).The COP 95%confidence ellipse area in the health group was the smallest,followed by the bilateral heel pain group,and finally the unilateral heel pain group,and the differences among 3 groups were statistically signifi-cant(P<0.05),there was also statistically significant difference between pairwise comparisons in 3 groups(P<0.05).Conclusion In the static condition,the pressure of the healthy heel and affected forefoot in the patients with PF is increased;while in the dynamic condition,the pressure of the toes of the healthy foot and forefoot of the affected foot in the patients with PF is also increased.The distribution of plantar stress in the patients with PF has larger difference compared with the healthy population,and the stability is poor.Meas-ures can be taken to improve the abnormal force on the foot,reduce pain and reduce the risk of falling.

This article summarizes Professor Huang Guicheng's clinical experience in the syndrome differentiation and treatment of lumbar spinal stenosis using the theory of"diseases of tendons,bones and collaterals".It is believed that lumbar spinal stenosis be-longs to the syndrome of deficiency in the root and excess in the superficiality,and is closely related to the three zang organs of the liv-er,spleen and kidney.Deficiency of the liver and kidney leading to a lack of the source of qi and blood,insufficiency of the spleen and stomach giving rise to phlegm and blood stasis,and the invasion of wind,cold and dampness pathogens cause the disease.The key to the pathogenesis lies in the obstruction of the collaterals.Taking the principles of"nourishing the collaterals and strengthening the healthy qi,dredging the collaterals and expelling the pathogens"as the therapeutic principles,Professor Huang is proficient in using insect drugs.The medicinal power can directly reach the collaterals,so that the collaterals can be unobstructed and the qi and blood can flow smoothly,achieving the therapeutic effect of expelling the pathogens,unblocking the collaterals and restoring the healthy qi.

HDAC7, a member of class IIa HDACs, plays a pivotal regulatory role in tumor, immune, fibrosis, and angiogenesis, rendering it a potential therapeutic target. Nevertheless, due to the high similarity in the enzyme active sites of class IIa HDACs, inhibitors encounter challenges in discerning differences among them. Furthermore, the substitution of key residue in the active pocket of class IIa HDACs renders them pseudo-enzymes, leading to a limited impact of enzymatic inhibitors on their function. In this study, proteolysis targeting chimera (PROTAC) technology was employed to develop HDAC7 drugs. We developed an exceedingly selective HDAC7 PROTAC degrader B14 which showcased superior inhibitory effects on cell proliferation compared to TMP269 in various diffuse large B cell lymphoma (DLBCL) and acute myeloid leukemia (AML) cells. Subsequent investigations unveiled that B14 disrupts BCL6 forming a transcriptional inhibition complex by degrading HDAC7, thereby exerting proliferative inhibition in DLBCL. Our study broadened the understanding of the non-enzymatic functions of HDAC7 and underscored the importance of HDAC7 in the treatment of hematologic malignancies, particularly in DLBCL and AML.

OBJECTIVE To analyze the blood components of Suanzaoren Decoction after oral administration using UHPLC-Q Exactive Orbitrap-MS technology.METHODS Female Sprague-Dawley(SD)rats were used as experimental subjects,and Suanza-oren Decoction was administered orally.Serum samples were collected,and the aqueous extract of Suanzaoren Decoction and the serum were analyzed using UHPLC-Q Exactive Orbitrap-MS technology to identify the prototype components and metabolites absorbed into the blood by comparing and analyzing with the LuMet-TCM database.RESULTS It showed that a total of 458 components were iden-tified in the aqueous extract of Suanzaoren Decoction,and 26 chemical components were identified in the blood,including 23 prototype components and 3 metabolites.CONCLUSION The prototype components absorbed into the blood discovered in this study may be the active ingredients of Suanzaoren Decoction,providing a reference for the research on the pharmacodynamic material basis of Suanza-oren Decoction.

This article summarizes Professor Huang Guicheng's clinical experience in the syndrome differentiation and treatment of lumbar spinal stenosis using the theory of"diseases of tendons,bones and collaterals".It is believed that lumbar spinal stenosis be-longs to the syndrome of deficiency in the root and excess in the superficiality,and is closely related to the three zang organs of the liv-er,spleen and kidney.Deficiency of the liver and kidney leading to a lack of the source of qi and blood,insufficiency of the spleen and stomach giving rise to phlegm and blood stasis,and the invasion of wind,cold and dampness pathogens cause the disease.The key to the pathogenesis lies in the obstruction of the collaterals.Taking the principles of"nourishing the collaterals and strengthening the healthy qi,dredging the collaterals and expelling the pathogens"as the therapeutic principles,Professor Huang is proficient in using insect drugs.The medicinal power can directly reach the collaterals,so that the collaterals can be unobstructed and the qi and blood can flow smoothly,achieving the therapeutic effect of expelling the pathogens,unblocking the collaterals and restoring the healthy qi.

OBJECTIVE To analyze the blood components of Suanzaoren Decoction after oral administration using UHPLC-Q Exactive Orbitrap-MS technology.METHODS Female Sprague-Dawley(SD)rats were used as experimental subjects,and Suanza-oren Decoction was administered orally.Serum samples were collected,and the aqueous extract of Suanzaoren Decoction and the serum were analyzed using UHPLC-Q Exactive Orbitrap-MS technology to identify the prototype components and metabolites absorbed into the blood by comparing and analyzing with the LuMet-TCM database.RESULTS It showed that a total of 458 components were iden-tified in the aqueous extract of Suanzaoren Decoction,and 26 chemical components were identified in the blood,including 23 prototype components and 3 metabolites.CONCLUSION The prototype components absorbed into the blood discovered in this study may be the active ingredients of Suanzaoren Decoction,providing a reference for the research on the pharmacodynamic material basis of Suanza-oren Decoction.

Tongue diagnosis is an important method of TCM diagnosis and treatment.Tongue is the key link of auxiliary diagnosis of tongue feature extraction and processing,and also is the bottleneck of intelligent tongue diagnosis in traditional Chinese medicine.Using image processing,artificial intelligence technology to the tongue as a quantitative and identify characteristics of traditional Chinese medicine,looking for both conforms to the original thinking of TCM,and TCM tongue diagnosis method of accurately,has become a common concern of traditional Chinese medicine and computer field.From the mobile terminal tongue as auxiliary diagnostic system of traditional Chinese medicine tongue acquisition basic attribute,tongue diagnosis and image information building,tongue like features are required for accurate extraction and so on related key technology is analyzed,and build overall architecture,so as to provide technical reference for the tongue like intelligent diagnosis,promote the development of technology of tongue diagnosis in traditional Chinese medicine modernization.

Objective:To explore the involvement of serum exosomal miR-17-5p in the pathogenesis of regulatory T (Treg) cells deficiency in premature ovarian insufficiency (POI) patients.Methods:In a case-control study, 32 Chinese Han women with POI and 32 Chinese Han women with regular menstrual cycles (healthy control, HC) attending the Department of Gynecological Endocrinology, the First Affiliated Hospital of Nanjing Medical University from January 2019 to December 2020 were recruited. The relative expressional level of serum exosomal miR-17-5p was confirmed by reverse transcription quantitative polymerase chain reaction (RT-qPCR). To understand how POI-exosome affects Treg cells, peripheral blood mononuclear cells (PBMC) of healthy volunteer were transfected with miR-17-5p, and cultured with PBS as the control or with exosomes derived from POI women or HC for 72 h. Subsequently, to evaluate the functionality of the miR-17-5p upregulated in POI-exosome, native CD4 + T cells from healthy donors were sorted by magnetic beads and induced into induced Treg (iTreg) cells in vitro, and the induction efficiency, the proliferation and apoptosis rates of iTreg cells were detected after transfection with miR-17-5p mimic/inhibitor. TargetScan, miRDB, miRTarBase and miRWalk were used to predict targets gene of miR-17-5p. Results:The expression level of serum exosomal miR-17-5p was markedly up-regulated in POI compared with HC ( P<0.001). Furthermore, after culture with POI-exosome [(0.93±0.40)%], the frequency of Treg cells was significantly reduced as compared with that culture with PBS [(3.77±0.89)%, P=0.005]. Subsequently, the frequency of Treg cells in miR-17-5p mimic group [(4.30±1.91)%] was significantly lower than that in negative control group [(11.97±1.82)%, P=0.007]. In contrast, both Th1 and Th17 did not be affected (both P>0.05). The induction efficiency of iTreg cells did not be influenced by transfection with miR-17-5p (both P>0.05), however, the iTreg cells apoptosis rate in miR-17-5p mimic group [(16.31±1.27)%] was significantly higher than that in negative control group [(13.01±1.80)%, P=0.035], while apoptosis rate was significantly lower [(7.97±0.71)%] and proliferation rate was higher [(88.83±0.25)%] in miR-17-5p inhibitor group than those in negative control group [(11.42±0.23)%, P=0.001; (87.60±0.70)%, P=0.045]. Transforming growth factor beta receptor 2 was most likely targets gene of miR-17-5p. Conclusion:The exosomal miRNA profile of patients with POI differed from that of HC, and miR-17-5p, which is markedly increased in POI patients, decreased the frequency of Treg cells by inhibiting iTreg cell proliferation and promoting iTreg cells apoptosis. Thus, our study suggested that exosomal miR-17-5p promoted iTreg cells apoptosis and resulted in Treg cells deficiency in POI pathogenesis.

Objective:To explore the involvement of serum exosomal miR-17-5p in the pathogenesis of regulatory T (Treg) cells deficiency in premature ovarian insufficiency (POI) patients.Methods:In a case-control study, 32 Chinese Han women with POI and 32 Chinese Han women with regular menstrual cycles (healthy control, HC) attending the Department of Gynecological Endocrinology, the First Affiliated Hospital of Nanjing Medical University from January 2019 to December 2020 were recruited. The relative expressional level of serum exosomal miR-17-5p was confirmed by reverse transcription quantitative polymerase chain reaction (RT-qPCR). To understand how POI-exosome affects Treg cells, peripheral blood mononuclear cells (PBMC) of healthy volunteer were transfected with miR-17-5p, and cultured with PBS as the control or with exosomes derived from POI women or HC for 72 h. Subsequently, to evaluate the functionality of the miR-17-5p upregulated in POI-exosome, native CD4 + T cells from healthy donors were sorted by magnetic beads and induced into induced Treg (iTreg) cells in vitro, and the induction efficiency, the proliferation and apoptosis rates of iTreg cells were detected after transfection with miR-17-5p mimic/inhibitor. TargetScan, miRDB, miRTarBase and miRWalk were used to predict targets gene of miR-17-5p. Results:The expression level of serum exosomal miR-17-5p was markedly up-regulated in POI compared with HC ( P<0.001). Furthermore, after culture with POI-exosome [(0.93±0.40)%], the frequency of Treg cells was significantly reduced as compared with that culture with PBS [(3.77±0.89)%, P=0.005]. Subsequently, the frequency of Treg cells in miR-17-5p mimic group [(4.30±1.91)%] was significantly lower than that in negative control group [(11.97±1.82)%, P=0.007]. In contrast, both Th1 and Th17 did not be affected (both P>0.05). The induction efficiency of iTreg cells did not be influenced by transfection with miR-17-5p (both P>0.05), however, the iTreg cells apoptosis rate in miR-17-5p mimic group [(16.31±1.27)%] was significantly higher than that in negative control group [(13.01±1.80)%, P=0.035], while apoptosis rate was significantly lower [(7.97±0.71)%] and proliferation rate was higher [(88.83±0.25)%] in miR-17-5p inhibitor group than those in negative control group [(11.42±0.23)%, P=0.001; (87.60±0.70)%, P=0.045]. Transforming growth factor beta receptor 2 was most likely targets gene of miR-17-5p. Conclusion:The exosomal miRNA profile of patients with POI differed from that of HC, and miR-17-5p, which is markedly increased in POI patients, decreased the frequency of Treg cells by inhibiting iTreg cell proliferation and promoting iTreg cells apoptosis. Thus, our study suggested that exosomal miR-17-5p promoted iTreg cells apoptosis and resulted in Treg cells deficiency in POI pathogenesis.