AIM:This study aims to investigate the effects of aerobic exercise on hypothalamic autophagy and central leptin resistance in obese mice,and to explore the potential mechanisms.METHODS:Forty male C57BL/6J mice,aged 7 to 8 weeks,were randomly assigned to 5 groups:normal control(CON)group,high-fat diet(HFD)group,HFD+exercise(HFD+Exe)group,HFD+phosphate-buffered saline(PBS)group,and HFD+rilmenidine(autophagy ago-nist)group,with 8 mice in each group.Additionally,twelve fibronectin type Ⅲ domain-containing protein 5(Fndc5)gene(encoding irisin precursor protein)knockout(Fndc5 KO)mice were randomly allocated to Fndc5 KO+HFD group and Fndc5 KO+HFD+Exe group,with 6 mice in each group.All mice were fed for 28 weeks.The mice in CON group re-ceived a normal diet,while those in the remaining groups were provided with an HFD.The mice in HFD+Exe and Fndc5 KO+HFD+Exe groups engaged in aerobic treadmill exercise while continuing an HFD from weeks 17 to 28.The mice in HFD+PBS group received intraperitoneal injections of PBS as a control,while those in HFD+rilmenidine group received in-traperitoneal injections of rilmenidine(10 mg?kg-1?d-1),4 times a week over a total duration of 12 weeks(weeks 17 to 28).Following the intervention,serum metabolite levels,as well as concentrations of leptin and irisin,were quantified by ELISA.Morphological alterations in the liver and white adipose tissues were evaluated through oil red O staining and he-matoxylin-eosin(HE)staining.Western blot was utilized to assess the hypothalamic protein levels of autophagy markers,autophagy-related protein 7(ATG7),beclin-1,microtubule-associated protein 1 light chain 3(LC3)and p62,and leptin resistance markers,suppressor of cytokine signaling 3(SOCS3)and protein tyrosine phosphatase 1B(PTP1B).RE-SULTS:Observations of mouse phenotypes indicated that HFD feeding significantly increased body weight,blood lipid content and serum leptin level(P<0.05).The results of HE and oil red O staining demonstrated that HFD feeding marked-ly promoted lipid accumulation in the liver and caused ballooning of white adipocytes.Western blot analyses revealed that HFD feeding significantly down-regulated the protein levels of ATG7,beclin-1 and LC3-Ⅱ/LC3-I,but up-regulated the pro-tein level of p62(P<0.05),thus reducing cellular autophagy capacity.Furthermore,HFD feeding elevated the protein levels of leptin resistance markers SOCS3 and PTP1B(P<0.05).Aerobic exercise and autophagy agonist were found to partially reverse these changes,enhancing cellular autophagy capacity and alleviating leptin resistance.However,these effects were diminished after knockout of Fndc5 gene,further substantiating the role of irisin in exercise-mediated en-hancement of cellular autophagy and attenuation of leptin resistance.CONCLUSION:Aerobic exercise alleviates hypo-thalamic autophagy defect and central leptin resistance in obese mice,which may be associated with exercise-induced irisin.

AIM:This study aims to investigate the effects of aerobic exercise on hypothalamic autophagy and central leptin resistance in obese mice,and to explore the potential mechanisms.METHODS:Forty male C57BL/6J mice,aged 7 to 8 weeks,were randomly assigned to 5 groups:normal control(CON)group,high-fat diet(HFD)group,HFD+exercise(HFD+Exe)group,HFD+phosphate-buffered saline(PBS)group,and HFD+rilmenidine(autophagy ago-nist)group,with 8 mice in each group.Additionally,twelve fibronectin type Ⅲ domain-containing protein 5(Fndc5)gene(encoding irisin precursor protein)knockout(Fndc5 KO)mice were randomly allocated to Fndc5 KO+HFD group and Fndc5 KO+HFD+Exe group,with 6 mice in each group.All mice were fed for 28 weeks.The mice in CON group re-ceived a normal diet,while those in the remaining groups were provided with an HFD.The mice in HFD+Exe and Fndc5 KO+HFD+Exe groups engaged in aerobic treadmill exercise while continuing an HFD from weeks 17 to 28.The mice in HFD+PBS group received intraperitoneal injections of PBS as a control,while those in HFD+rilmenidine group received in-traperitoneal injections of rilmenidine(10 mg?kg-1?d-1),4 times a week over a total duration of 12 weeks(weeks 17 to 28).Following the intervention,serum metabolite levels,as well as concentrations of leptin and irisin,were quantified by ELISA.Morphological alterations in the liver and white adipose tissues were evaluated through oil red O staining and he-matoxylin-eosin(HE)staining.Western blot was utilized to assess the hypothalamic protein levels of autophagy markers,autophagy-related protein 7(ATG7),beclin-1,microtubule-associated protein 1 light chain 3(LC3)and p62,and leptin resistance markers,suppressor of cytokine signaling 3(SOCS3)and protein tyrosine phosphatase 1B(PTP1B).RE-SULTS:Observations of mouse phenotypes indicated that HFD feeding significantly increased body weight,blood lipid content and serum leptin level(P<0.05).The results of HE and oil red O staining demonstrated that HFD feeding marked-ly promoted lipid accumulation in the liver and caused ballooning of white adipocytes.Western blot analyses revealed that HFD feeding significantly down-regulated the protein levels of ATG7,beclin-1 and LC3-Ⅱ/LC3-I,but up-regulated the pro-tein level of p62(P<0.05),thus reducing cellular autophagy capacity.Furthermore,HFD feeding elevated the protein levels of leptin resistance markers SOCS3 and PTP1B(P<0.05).Aerobic exercise and autophagy agonist were found to partially reverse these changes,enhancing cellular autophagy capacity and alleviating leptin resistance.However,these effects were diminished after knockout of Fndc5 gene,further substantiating the role of irisin in exercise-mediated en-hancement of cellular autophagy and attenuation of leptin resistance.CONCLUSION:Aerobic exercise alleviates hypo-thalamic autophagy defect and central leptin resistance in obese mice,which may be associated with exercise-induced irisin.

Abstract: The differential expression and subcellular localization of single-cell proteins are closely related to the physiological state and pathological mechanisms of the body. The development of single-cell protein in situ imaging methods provides powerful tools for spatial single-cell proteomics research and single-cell protein profiling. This article summarizes the single-cell protein imaging methods developed in recent years, including the circulating immunofluorescence imaging methods based on ordered multi-round antibody incubation, mass spectrometry imaging based on metal element labeled antibodies, fluorescence imaging based on DNA-barcoded antibody, gene encoded fluorescence protein imaging and spectral imaging based on Raman spectroscopy or X-ray spectroscopy, with brief explanation of the imaging principles of these methods. It focuses on the multiple performance, imaging resolution and signal amplification performance of these methods, and analyzes their application characteristics in practical scientific research and clinical work, in the hope of providing some reference for the development of more revolutionary single-cell imaging methods, and promoting the development of biomedical and precision medicine.

BACKGROUND:Muscle weakness is a common symptom after coronavirus disease 2019(COVID-19)infection and affects the ability to perform daily activities in humans during recovery.Low-frequency pulsed magnetic field stimulation at a strength of 1.5 mT and a frequency of 3 300 Hz can enhance the maximal voluntary contraction and strength endurance of human skeletal muscle by inducing and activating classical transient receptor potential channel 1(TRPC1),which produces a series of pathological support effects on muscle tissue.It has not been studied whether this means will improve muscle weakness in patients recovering from COVID-19. OBJECTIVE:To select the low-frequency pulsed magnetic field for magnetic stimulation of lower limb muscle groups in patients with COVID-19,in order to observe the effect of this stimulation on the improvement of muscle weakness of lower limb muscle groups in patients with COVID-19 during the recovery period. METHODS:Fourteen patients infected with COVID-19(Omicron strain)positive for Innovita COVID-19 Ab Test(Colloidal Gold)and accompanied by muscle weakness were recruited and randomly divided into two groups:a test group receiving magnetic field stimulation and a control group receiving sham treatment,respectively.The total duration of the trial was 3 weeks.The test group was given low-frequency pulsed magnetic stimulation of the lower limbs every 48 hours and the control group was given the same intervention procedure as the test group but with sham stimulation.Patients in both groups were not informed whether the magnetic stimulation apparatus was running or not.Nine sessions were performed in both groups and the changes in the maximum voluntary contraction,explosive leg force and strength endurance of the local muscle groups of the lower limbs were subsequently observed in both groups. RESULTS AND CONCLUSION:Among the eight local muscle groups collected,seven local muscle groups in the test group showed an increase in the maximum voluntary contraction value after 3 weeks of low-frequency pulsed magnetic field stimulation.In the control group,there were only three muscle groups with improvement in the maximum voluntary contraction.The rate of improvement in the anterior and posterior muscle groups of the left leg in the test group was significantly higher than that in the control group.The longitudinal jump height and peak angular velocity of the knee joint in both groups were improved compared with the pre-test measurement,and the elevation rate of jumping height in the test group was higher than that in the control group.Under the fatigue condition,the decline rates of peak angular velocity of the knee joint and jumping height in the test group decreased significantly,while those in the control group did not change significantly.The above data confirmed that the low-frequency pulsed magnetic field stimulation with the intensity of 1.5 mT and frequency of 3 300 Hz could improve the muscle strength of more local muscle groups in the lower limbs of patients with COVID-19 during the recovery period compared with the human self-healing process,and the whole-body coordination ability and functional status based on explosive leg force of the legs could be significantly improved.Therefore,low-frequency pulsed magnetic field stimulation can be used as an effective,non-exercise rehabilitation tool to improve muscle weakness in the lower limbs of patients with COVID-19.

Osteoporosis can be induced by various factors including prolonged glucocorticoid usage, diminished estrogen levels, secondary hyperparathyroidism, and alterations in the microenvironment of bone tissue. The bone metabolism imbalance(osteogenic-lipogenic imbalance) plays a crucial role in the development of osteoporosis. This imbalance is primarily driven by an increase in the differentiation of bone marrow mesenchymal stem cells into adipocytes and a decrease in their differentiation into osteoblasts, thus forming the core of the osteogenic-lipogenic imbalance observed in osteoporosis. The bone morphogenesis protein(BMP) plays a crucial role in the regulation of the osteogenic-lipid balance in osteoporosis. This regulatory function is accomplished through both the Smad-dependent and Smad-independent pathways. This review centers on the Smad-dependent and Smad-independent pathways facilitated by BMP, offering a comprehensive overview of the potential mechanisms through which BMP-2, 4, 6, 7, and 9 contribute to the regulation of osteogenesis and lipid metabolism in osteoporosis via these pathways. In order to present novel insights for the identification of efficacious targets for clinical anti-osteoporosis medications.

Osteoporosis(OP)is a chronic systemic metabolic bone disease.In recent years,with development of population aging,incidence of OP is also increasing,bringing huge economic burden to family and society.There are many causes of OP,such as aging,oxidative stress,epigenetic and many other aspects,and with further development of research,inflammatory factors and other bone immunology into people's vision.Th17 cells are effector T cell subpopulation that induce development of inflammation and promote bone loss.On the contrary,Treg can maintain their own tolerance and further suppress their own immunity,thus playing a bone protective role.Under physiological conditions,Th17/Treg balance can maintain bone homeostasis,while under pathological condi-tions,disruption of Th17/Treg balance can lead to bone loss.Based on this,this paper will review pathways and cytokines mediating Th17/Treg balance in OP,in order to provide new ideas and methods for treatment of OP.

Non-coding RNAs(ncRNAs) are special RNAs that they don't have protein coding function, but they can affect chromosome structure, gene transcription and participate in the processes of epigenetic modifications. ncRNAs include long non-coding RNAs, microRNA, etc. In recent years, it has been found that these ncRNAs can maintain bone remodeling by adjusting bone resorption and formation in osteoporosis(OP). In the future, it may be a key target of the drug action screening which is clarifying the regulatory mechanism of ncRNAs in the occurrence and development of OP. OP belongs to bone rheumatism category in traditional Chinese medicine, according to the theory of “the kidney generating marrow and dominating bone” in traditional Chinese medicine, kidney tonifying and bone strengthening formulas are used to treat the OP in clinic, and the curative effect is remarkable. It has been found that kidney tonifying and bone strengthening prescriptions can enhance the proliferation of osteoblasts or inhibit the differentiation of osteoclasts by up-regulating or down-regulating the expression of ncRNA, and finally maintain OP bone homeostasis, thus exerting therapeutic effect. However, the specific molecular mechanism is still in its exploratory stage. Therefore, this paper summarized the molecular mechanism of kidney tonifying and bone strengthening prescriptions regulating ncRNAs in the treatment of OP in recent years, in order to provide the new ideas for the screening of the key therapeutic targets of OP drugs and the prevention and treatment of OP with traditional Chinese medicine.

Objective:To quantify the plasma concentrations of inorganic arsenic (As(III) and As(V)) and methylated metabo-lites ( MMA and DMA) , and to detect the total amount of arsenic in blood cells and plasma by high performance liquid chromatogra -phy-hydridegeneration-atomic fluorescence spectrometry ( HPLC-HG-AFS) and HG-AFS methods to clarify the arsenic species in acute promyelocytic leukemia (APL) patients.Methods:The blood cells and plasma were digested by the mixture of HNO 3-H2O2 and ana-lyzed by HG-AFS.For the arsenic species , the plasma samples were prepared with perchloric acid to precipitate protein .The superna-tant was separated on an anion-exchange column in 6 min with isocratic elution using 13 mmol · L-1 CH3 COONa, 3 mmol · L-1 NaH2 PO4 , 4 mmol· L-1 KNO3 and 0.2 mmol· L-1 EDTA-2Na.Results:The methods provided linear range of 0.2-20 ng· ml-1 for total arsenic and 2.0-50 ng· ml-1 for four arsenic species (r>0.9950).The spiked recoveries ranged from 81.2％to 108.6％, and the coefficients of variation for intra-and inter-batch precision were less than 9.3％and 12.5％, respectively.The developed methods were applied successfully in the assay of total arsenic and arsenic species in 5 APL patients.Conclusion:The method is simple, fast and accurate , which can be applied in the assay of arsenic compounds in plasma and blood cells in APL patients .

Objective To compare biliary tract intraductal papillary mucinous neoplasm (BT-IPMN) with intraductal papillary mucinous neoplasm of the pancreas (P-IPMN).Methods From January 2008 to December 2013,eleven (6.0％) cases of BT-IPMNs were retrospectively identified from a total of 182 biliary tract tumors resected in our institution,while 50 cases underwent surgery for P-IPMN.The mean age of the 11 BT-IPMN cases was 57.3 years (range 40 to 74 years).There were 8 men (73.0％).The mean age of the 50 P-IPMN cases was 57.4 years (range 33 to 85 years).There were 34 men (68.0％).The clinical features,radiologic findings,pathology,surgical strategies,and long-term follow up outcomes between the 2 groups of patients were analyzed.Results There was no significant difference between BT-IPMN and P-IPMN in the following aspects:mean age,sex ratios and clinical presentation [including the most common presenting symptom abdominal pain (73.0％ vs.68.0％),and elevated tumor markers (CEA and CA19-9)].The mean tumor size of BT-IPMN was significantly smaller than P-IPMN (1.7 vs.4.1 cm,P ＜ 0.05).Macroscopically visible mucin was detected in all the 11 patients basing on the original surgical reports.The most common abnormal preoperative imaging findings for BT-IPMN were bile duct dilatation (100％) and intraluminal masses (55.0％).Most cases (82.0％) involved the intrahepatic bile duct and hilum.For tumor clearance,we conducted left hepatectomy in most cases (64.0％).Only one patient underwent biopsy and choledochojejunostomy for multiple tumors involving the extrahepatic,right and left bile ducts.BT-IPMN was likely to have a higher risk of malignancy (55.0％ vs.44.0％) and poorer prognosis (median survival,57 vs.63 months),although there was no significant difference (P ＞ 0.05).The patient without tumor resection died of liver failure 22 months after palliative surgery.Conclusions BT-IPMNs are very rare and they had some similarity with P-IPMN.Complete resection of BT-IPMN is associated with good survival.

BACKGROUND:The effects of engineered bone scaffold containing seeding cels with different shapes to repair bone defect are varied, while the loaded cellquantity is the important factor influencing the curative effect, but which is rarely reported. OBJECTIVE:By preparing self-made corrugated tissue-engineered bone scaffold and other three forms of bone tissue engineering scaffolds, to study the quantity of loaded cels on different scaffolds and osteogenesis of corrugated tissue-engineered bone scaffold so as to discuss the advantages and features of self-made corrugated tissue-engineered bone scaffold. METHODS: (1) Experimentin vitro: There were four kinds of scaffolds with the same volume and samples: calcium phosphate cement (CPC) corrugated surface scaffold group, smooth surface scaffold group, cylindrical scaffold group and porous cylindrical scaffold with holow tubes group, in which the latter three groups are control ones. A certain volume with same density of rabbit bone marrow mesenchymal stem cels (BMSCs) suspension after osteogenesis induction was seeded onto the scaffolds. After incubation, culture, digestion and colection, cellquantity was counted, absorbance value was finaly detected and cellactivity was proofed by alkaline phosphatase and alizarin red staining. (2) Experimentin vivo: New Zealand rabbits were randomly and equaly divided into recombinant human bone morphogenetic protein-2 (rhBMP-2)/CPC/BMSCs corrugated scaffold group, pure CPC corrugated scaffold group and cancelous bone implant group. Three kinds of scaffold implants with the same volume were inserted into the area between rabbit’s L5, 6 transverse processes bilateraly. At 4, 8, 12 weeks postoperatively, gross and histological observation was performed. RESULTS AND CONCLUSION:(1)Experimentin vitro: The drip of cellsuspension steadily stayed on the surface of corrugated scaffold because of corrugated shape groove and the surface tension of the liquid. The amount of cels per sample digested down from the CPC corrugated surface scaffold was significantly more than that from the other three groups (P < 0.05), while the absorbance values did not differ (P > 0.05). (2) Experimentin vivo: At each time point the osteogenesis quantity of rhBMP-2/CPC/BMSCs corrugated scaffold group was more than that of the pure CPC corrugated scaffold group (P < 0.05), while there was no difference from the cancelous bone implant group (P > 0.05). These findings indicate that the characteristics of the self-made corrugated engineered bone scaffold are beneficial to seed cellloading, which supports a large number of osteogenesis and provides feasibility to promote the healing of segmental bone defects.