Objective:To establish the UPLC fingerprint evaluation system of Viticis Fructus; To comprehensively evaluate the quality of Viticis Fructus and its processed products combining with multivariate statistical methods and compositional variance analysis.Methods:19 batches of Viticis Fructus from different regions were collected and processed by frying process into decoction pieces. The separation was operated on Waters CORTECS T3 C18 chromatographic column (100 mm × 2.1 mm, 1.6 μm). Acetonitrile and 0.1% phosphoric acid water were used as mobile phases for gradient elution, to establish the UPLC fingerprints of Viticis Fructus. The UPLC fingerprints of Viticis Fructus were analyzed using similarity evaluation, principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA). The contents of seven active components in the samples of Viticis Fructus and fried Viticis Fructus were determined.Results:A total of 26 common peaks were identified in the fingerprints of 38 batches of samples, and 7 components were identified. Similarity evaluation results demonstrated that the chemical components of Viticis Fructus from Jingdezhen City, Jiangxi Province, were significantly different from those of other regions. The results of PCA and PLS-DA analysis showed that the chemical components of Viticis Fructus and fried Viticis Fructus could be clearly distinguished, and the processing process had an impact on the components. The results of content determination showed that the contents of some components increased or decreased after frying. The analysis results of grey correlation method and TOPSIS method show that the medicinal materials in Jiujiang City, Jiangxi Province have a high score ranking and stable quality.Conclusion:This study successfully establishes the fingerprints of Viticis Fructus and its processed products, grey correlation method and TOPSIS method analysis revealed the quality differences of samples from different origins, which providing a scientific basis for the quality control and evaluation of Viticis Fructus and its processed products.

Objective To explore the mechanism of electroacupuncture in alleviating hyperlipidemia in a rat model by modulating mammalian target of rapamycin complex 1(mTORC1)-mediated lipophagy in hepatocytes.Methods A total of 30 SD rats were randomly divided into blank(n=6)and modeling groups(n=24)using the random number table method.A hyperlipidemic rat model was established by feeding rats a high-fat diet(feeding for 8 weeks).After successful modeling,the modeling group was randomly divided into the model,electroacupuncture,mTORC1 inhibitor,and electroacupuncture+mTORC1 agonist groups,with six rats in each group.Except for the blank group,all other rats were fed with high fat diet.Rats in the electroacupuncture and electroacupuncture+mTORC1 agonist groups received electroacupuncture intervention at bilateral"Fenglong"(ST40)acupoints(dilatational wave 2 Hz/100 Hz,current intensity 1 mA)for 30 min once daily.Rats in the mTORC1 inhibitor group received intraperitoneal injections of the mTORC1 inhibitor,rapamycin(2 mg/kg),once daily.Rats in the electroacupuncture+mTORC1 agonist group received intraperitoneal injections of the mTORC1 agonist MHY1485(10 mg/kg)once daily.The interventions were administered for five consecutive days per week for 4 weeks.Upon completion of the intervention,the following analyses were performed:serum contents of total cholesterol(TC),triglycerides(TAG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),free fatty acids(FFA),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were measured using a fully automated biochemical analyzer.Hepatic histopathological changes and lipid deposition were observed using hematoxylin-eosin and oil red O staining.The liver condition was observed and the liver index was calculated.Hepatic TC and TAG levels were measured using an enzyme-linked immunosorbent assay.The ultrastructure of the liver tissue was observed using transmission electron microscopy,and the mean fluorescence intensity of perilipin 2(PLIN2)and microtubule-associated protein 1 light chain 3(LC3)-Ⅱ in the liver tissue was detected using immunofluorescence.Protein expression of LC3-Ⅱ/LC3-Ⅰ,phosphorylated mammalian target of rapamycin(p-mTOR)/mTOR,and mTORC1 in liver tissue was detected using Western blotting.Results Compared to the blank group,the model group rats showed increased serum TC,TAG,LDL-C,ALT,AST,and FFA levels,along with decreased HDL-C levels(P<0.05).The liver index and hepatic TC and TAG levels were also elevated(P<0.05).Histological examination of liver tissue revealed substantial lipid accumulation,numerous lipid droplets within hepatocytes,abnormal mitochondrial morphology,and scarce autophagic vacuole.The mean fluorescence intensity of PLIN2 increased,whereas that of LC3-Ⅱ decreased(P<0.05).Additionally,the LC3-Ⅱ/LC3-Ⅰ ratio was reduced,whereas the p-mTOR/mTOR ratio and mTORC1 protein expression were increased(P<0.05).Compared to the model group,rats in the mTORC1 inhibitor and electroacupuncture groups exhibited decreased serum TC,TAG,LDL-C,ALT,AST,and FFA levels(P<0.05),along with a reduced liver index and hepatic TC and TAG levels(P<0.05).Histological examination showed markedly attenuated lipid accumulation and visible autophagic vacuole in the hepatocytes.The mean fluorescence intensity of PLIN2 decreased,whereas that of LC3-Ⅱ increased(P<0.05).Moreover,the LC3-Ⅱ/LC3-Ⅰ ratio increased,whereas the p-mTOR/mTOR ratio and mTORC1 protein expression decreased(P<0.05).In comparison with both the electroacupuncture and mTORC1 inhibitor groups,the electroacupuncture+mTORC1 agonist group demonstrated increased serum TAG,TC,LDL-C,ALT,AST,and FFA levels(P<0.05)as well as elevated liver index and hepatic TC and TAG levels(P<0.05).Liver tissues exhibited aggravated lipid deposition and absence of autophagic vacuole in liver cells.The mean fluorescence intensity of PLIN2 was enhanced,whereas that of LC3-Ⅱ was reduced(P<0.05).Furthermore,the LC3-Ⅱ/LC3-Ⅰ ratio decreased,and the p-mTOR/mTOR ratio and mTORC1 protein expression increased(P<0.05).Conclusion Electroacupuncture at"Fenglong"(ST40)may improve blood lipid levels in hyperlipidemic rats by inhibiting mTORC1 and promoting hepatocyte lipophagy.

Objective To explore the mechanism of electroacupuncture in alleviating hyperlipidemia in a rat model by modulating mammalian target of rapamycin complex 1(mTORC1)-mediated lipophagy in hepatocytes.Methods A total of 30 SD rats were randomly divided into blank(n=6)and modeling groups(n=24)using the random number table method.A hyperlipidemic rat model was established by feeding rats a high-fat diet(feeding for 8 weeks).After successful modeling,the modeling group was randomly divided into the model,electroacupuncture,mTORC1 inhibitor,and electroacupuncture+mTORC1 agonist groups,with six rats in each group.Except for the blank group,all other rats were fed with high fat diet.Rats in the electroacupuncture and electroacupuncture+mTORC1 agonist groups received electroacupuncture intervention at bilateral"Fenglong"(ST40)acupoints(dilatational wave 2 Hz/100 Hz,current intensity 1 mA)for 30 min once daily.Rats in the mTORC1 inhibitor group received intraperitoneal injections of the mTORC1 inhibitor,rapamycin(2 mg/kg),once daily.Rats in the electroacupuncture+mTORC1 agonist group received intraperitoneal injections of the mTORC1 agonist MHY1485(10 mg/kg)once daily.The interventions were administered for five consecutive days per week for 4 weeks.Upon completion of the intervention,the following analyses were performed:serum contents of total cholesterol(TC),triglycerides(TAG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),free fatty acids(FFA),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were measured using a fully automated biochemical analyzer.Hepatic histopathological changes and lipid deposition were observed using hematoxylin-eosin and oil red O staining.The liver condition was observed and the liver index was calculated.Hepatic TC and TAG levels were measured using an enzyme-linked immunosorbent assay.The ultrastructure of the liver tissue was observed using transmission electron microscopy,and the mean fluorescence intensity of perilipin 2(PLIN2)and microtubule-associated protein 1 light chain 3(LC3)-Ⅱ in the liver tissue was detected using immunofluorescence.Protein expression of LC3-Ⅱ/LC3-Ⅰ,phosphorylated mammalian target of rapamycin(p-mTOR)/mTOR,and mTORC1 in liver tissue was detected using Western blotting.Results Compared to the blank group,the model group rats showed increased serum TC,TAG,LDL-C,ALT,AST,and FFA levels,along with decreased HDL-C levels(P<0.05).The liver index and hepatic TC and TAG levels were also elevated(P<0.05).Histological examination of liver tissue revealed substantial lipid accumulation,numerous lipid droplets within hepatocytes,abnormal mitochondrial morphology,and scarce autophagic vacuole.The mean fluorescence intensity of PLIN2 increased,whereas that of LC3-Ⅱ decreased(P<0.05).Additionally,the LC3-Ⅱ/LC3-Ⅰ ratio was reduced,whereas the p-mTOR/mTOR ratio and mTORC1 protein expression were increased(P<0.05).Compared to the model group,rats in the mTORC1 inhibitor and electroacupuncture groups exhibited decreased serum TC,TAG,LDL-C,ALT,AST,and FFA levels(P<0.05),along with a reduced liver index and hepatic TC and TAG levels(P<0.05).Histological examination showed markedly attenuated lipid accumulation and visible autophagic vacuole in the hepatocytes.The mean fluorescence intensity of PLIN2 decreased,whereas that of LC3-Ⅱ increased(P<0.05).Moreover,the LC3-Ⅱ/LC3-Ⅰ ratio increased,whereas the p-mTOR/mTOR ratio and mTORC1 protein expression decreased(P<0.05).In comparison with both the electroacupuncture and mTORC1 inhibitor groups,the electroacupuncture+mTORC1 agonist group demonstrated increased serum TAG,TC,LDL-C,ALT,AST,and FFA levels(P<0.05)as well as elevated liver index and hepatic TC and TAG levels(P<0.05).Liver tissues exhibited aggravated lipid deposition and absence of autophagic vacuole in liver cells.The mean fluorescence intensity of PLIN2 was enhanced,whereas that of LC3-Ⅱ was reduced(P<0.05).Furthermore,the LC3-Ⅱ/LC3-Ⅰ ratio decreased,and the p-mTOR/mTOR ratio and mTORC1 protein expression increased(P<0.05).Conclusion Electroacupuncture at"Fenglong"(ST40)may improve blood lipid levels in hyperlipidemic rats by inhibiting mTORC1 and promoting hepatocyte lipophagy.

Objective To establish a rat model of type 2 diabetes mellitus with hepatic fibrosis using a high-lipid-sugar diet(HLSD)combined with inducement with carbon tetrachloride(CCl4)and streptozotocin.Methods SD rats were randomly divided into a liver fibrosis(LF)group,type 2 diabetes combined with liver fibrosis(DLF)group,and normal group,with 10 rats in each group.All rats in the LF or DLE group were fed an HLSD for 4 weeks;then,all rats were intraperitoneally injected with 20％CCl4(once a week)according to body weight(5 mL/kg)to induce hepatic fibrosis for another 4 weeks.During CCl4 treatment,rats in the DLF group were also intraperitoneally injected with 25 mg/kg STZ(once a week).After STZ treatment,the contents of FBG,INS,HOMA-IR,and HbA1c were evaluated once a week.The level of FBG was detected by blood glucose meter,and INS and HbA1c contents were detected by ELISA.Biochemical analysis was used to analyze the serum levels of AST,ALT,Tbil,and HA,as well as the HYP content in liver tissues.HE and Masson staining were used to evaluate pathological changes in liver tissues.Results The contents of FBG,HbA1c,and HOMA-IR were significantly increased,while the INS content was significantly decreased in the DLF group compared with those in the normal or LF group.Compared with the normal group,the LF group and DLF group showed higher levels of ALT,AST,Tbil,HA,and HYP,as well as fibrotic changes and inflammation in the liver tissue.Conclusions HLSD combined with CCl4 and STZ inducement is an effective strategy for establishing a rat model of type 2 diabetes mellitus with liver fibrosis.

Objective To establish a rat model of type 2 diabetes mellitus with hepatic fibrosis using a high-lipid-sugar diet(HLSD)combined with inducement with carbon tetrachloride(CCl4)and streptozotocin.Methods SD rats were randomly divided into a liver fibrosis(LF)group,type 2 diabetes combined with liver fibrosis(DLF)group,and normal group,with 10 rats in each group.All rats in the LF or DLE group were fed an HLSD for 4 weeks;then,all rats were intraperitoneally injected with 20％CCl4(once a week)according to body weight(5 mL/kg)to induce hepatic fibrosis for another 4 weeks.During CCl4 treatment,rats in the DLF group were also intraperitoneally injected with 25 mg/kg STZ(once a week).After STZ treatment,the contents of FBG,INS,HOMA-IR,and HbA1c were evaluated once a week.The level of FBG was detected by blood glucose meter,and INS and HbA1c contents were detected by ELISA.Biochemical analysis was used to analyze the serum levels of AST,ALT,Tbil,and HA,as well as the HYP content in liver tissues.HE and Masson staining were used to evaluate pathological changes in liver tissues.Results The contents of FBG,HbA1c,and HOMA-IR were significantly increased,while the INS content was significantly decreased in the DLF group compared with those in the normal or LF group.Compared with the normal group,the LF group and DLF group showed higher levels of ALT,AST,Tbil,HA,and HYP,as well as fibrotic changes and inflammation in the liver tissue.Conclusions HLSD combined with CCl4 and STZ inducement is an effective strategy for establishing a rat model of type 2 diabetes mellitus with liver fibrosis.

Objective This study aimed to evaluate the bone integration performance of antibacterial carbon dot(CD)-modified polyether ether ketone(PEEK)in infectious bone defect environments.Methods Guanidine-based CDs(G-CDs)prepared by the melting method combined with dialysis purification were used to modify PEEK implants using polyvinyl butyraldehyde(PVB)by the soaking-drying method(PEEK/PVB-G-CDs).SD rats were divided into the following groups:(1)PEEK-implanted uninfected(PEEK(-)),(2)PEEK/PVB-G-CDs-implanted uninfected(PEEK/PVB-G-CDs(-)),(3)PEEK-implanted infected(PEEK(+)),and(4)PEEK/PVB-G-CDs-implanted infected(PEEK/PVB-G-CDs(+)).A hole(diameter 2 mm,depth 5 mm)was drilled at the lateral condyle of the vertical femur in all rats to simulate a bone defect.Rats in the PEEK(-)and PEEK/PVB-G-CDs(-)groups without infection were injected with 30 μL physiological saline into the bone marrow cavity,and rats in the PEEK(+)and PEEK/PVB-G-CDs(+)groups with infection were injected with 30 μL MRSA bacterial suspension(1.5 × 104 colony-forming units/mL)into the bone marrow cavity.The implantation site was observed using animal-specific X-ray examination at 0,2,and 4 weeks after implantation,and the bone tissue characteristics of the implantation site were evaluated by micro computed tomography(CT)at 6 weeks after surgery.The bone implantation sites in each group of rats were examined by bacterial culture of bone marrow and hematoxylin and eosin staining,Toluidine blue,Goldner trichrome,and immunohistochemical staining.Results X-ray,Micro-CT,bacterial culture of bone marrow,and histopathological analysis confirmed no signs of infection in the PEEK(-)and PEEK/PVB-G-CDs(-)groups and the implants were integrated with the bone defects.Rats in the PEEK/PVB-G-CDs(+)group showed signs of antibacterial activity that effectively controlled the osteomyelitis caused by MRSA and achieved bone integration,while rats in the PEEK(+)group failed to achieve bone integration because of persistent infection.Immunohistochemical staining confirmed lower levels of anti-inflammatory factors such as IL-4 and IL-10 in the PEEK(+)group,and stronger expression of pro-inflammatory factors such as IL-6 and TNF-α compared with the other three groups,indicating that G-CD-modified PEEK inhibited MRSA infection,regulated inflammation levels in the local microenvironment,and promoted bone integration at the site of bone defects.Conclusions Antibacterial G-CDs modified PEEK exhibits excellent bone integration performance,providing a candidate strategy for future clinical treatment of infectious bone defects.

Objective This study aimed to evaluate the bone integration performance of antibacterial carbon dot(CD)-modified polyether ether ketone(PEEK)in infectious bone defect environments.Methods Guanidine-based CDs(G-CDs)prepared by the melting method combined with dialysis purification were used to modify PEEK implants using polyvinyl butyraldehyde(PVB)by the soaking-drying method(PEEK/PVB-G-CDs).SD rats were divided into the following groups:(1)PEEK-implanted uninfected(PEEK(-)),(2)PEEK/PVB-G-CDs-implanted uninfected(PEEK/PVB-G-CDs(-)),(3)PEEK-implanted infected(PEEK(+)),and(4)PEEK/PVB-G-CDs-implanted infected(PEEK/PVB-G-CDs(+)).A hole(diameter 2 mm,depth 5 mm)was drilled at the lateral condyle of the vertical femur in all rats to simulate a bone defect.Rats in the PEEK(-)and PEEK/PVB-G-CDs(-)groups without infection were injected with 30 μL physiological saline into the bone marrow cavity,and rats in the PEEK(+)and PEEK/PVB-G-CDs(+)groups with infection were injected with 30 μL MRSA bacterial suspension(1.5 × 104 colony-forming units/mL)into the bone marrow cavity.The implantation site was observed using animal-specific X-ray examination at 0,2,and 4 weeks after implantation,and the bone tissue characteristics of the implantation site were evaluated by micro computed tomography(CT)at 6 weeks after surgery.The bone implantation sites in each group of rats were examined by bacterial culture of bone marrow and hematoxylin and eosin staining,Toluidine blue,Goldner trichrome,and immunohistochemical staining.Results X-ray,Micro-CT,bacterial culture of bone marrow,and histopathological analysis confirmed no signs of infection in the PEEK(-)and PEEK/PVB-G-CDs(-)groups and the implants were integrated with the bone defects.Rats in the PEEK/PVB-G-CDs(+)group showed signs of antibacterial activity that effectively controlled the osteomyelitis caused by MRSA and achieved bone integration,while rats in the PEEK(+)group failed to achieve bone integration because of persistent infection.Immunohistochemical staining confirmed lower levels of anti-inflammatory factors such as IL-4 and IL-10 in the PEEK(+)group,and stronger expression of pro-inflammatory factors such as IL-6 and TNF-α compared with the other three groups,indicating that G-CD-modified PEEK inhibited MRSA infection,regulated inflammation levels in the local microenvironment,and promoted bone integration at the site of bone defects.Conclusions Antibacterial G-CDs modified PEEK exhibits excellent bone integration performance,providing a candidate strategy for future clinical treatment of infectious bone defects.

Objective: To study the effect of continuous static pressure on the endoplasmic reticulum of human periodontal ligament cells (hPDLCs) and the mechanism of osteogenic differentiation. Methods: hPDLCs cultured in vitro were subjected to 1 g/cm 2 of continuous compressive pressure (CCP) by custom-made, round, glass panes for 0, 2, 4, and 6 h, respectively. Alkaline phosphatase staining was used to detect osteogenic differentiation, and real-time quantitative PCR was used to detect the expression of protein kinase receptor-like ER kinase (PERK), eukaryotic translation initiation factor 2α (eIF2α), and transcription activation factor 4 (ATF-4). The 0 h loading group was the control group. Results: After CCP treatment, the alkaline phosphatase staining of hPDLCs was blue-violet and significantly stronger than that of cells in the control group. The expression levels of PERK and ATF4 in the hPDLCs after CCP treatment were higher than those of cells in the control group (P < 0.05) and increased over time (P < 0.05). The expression of eIF2α was lower in the experimental groups than in the control group (P < 0.05) and decreased over time (P < 0.05). Conclusion Mechanical stimulation can activate ERS in hPDLCs, leading to enhanced PERK-eIF2α-ATF4 signaling and inducing osteogenic differentiation.

Objective To study the effect of combined atorvastatin and ezetimibe pretreatment on perioperative hs-CRP after elective PCI.Methods One hundred and fifty-six patients with typical chronic stable angina pectoris were randomly divided into atorvastatin treatment group (n=78) and combined atorvastatin and ezetimibe treatment group (n=78).Their serum hs-CRP,TC and LDL-C level was measured before PCI,at hours 8,24,48 and on day 7 after PCI.Results In comparision with pre-operation,the serum TC and LDL-C levels were significantly lower in two groups (P＜0.01) and in combined atorvastatin and ezetimibe treatment group than in atorvastatin treatment group on day 7 after PCI (P＜0.05).The serum hs-CRP level was significantly higher in two groups at 8 h after PCI than before PCI,reached its peak at 24 h after PCI,continued to increase at 48 h after PCI (P＜0.01),no significant difference was found between the two groups on day 7 after PCI (P＞0.05).The average serum hs-CRP level was lower in combined atorvastatin and ezetimibe treatment group than in atorvastatin treatment group at hours 8,24 and 48 after PCI (P＜0.05) with no significant change found between the two groups on day 7 after PCI (P＞0.05).Conclusion The effect of combined atorvastatin and ezetimibe pretreatment is better than that of atorvastatin alone on perioperative acute inflammatory reactions after PCI.

Objective To investigate the characteristics of amygdala neural circuitry in comorbidity of late-life depression (LLD) and cognitive impairment. Methods Twenty-four LLD,eighteen amnestic mild cognitive impairments (aMCI),thirteen aMCI with depression (dMCI) and thirty cognitive normal (CN) subjects completed resting-state functional magnetic resonance imaging scan. Main effects of depression and MCI and their interactions on the intrinsic amygdala functional connectivity network ( AFCN) connectivity were examined. Behavioral significance of AFCN that voxel-wised amygdala connectivity correlating with de-pression severity and memory scores were also tested after controlling the effects of covariates,including age, gender,education, gray matter atrophy, and group. Results The immediate memory and delayed memory function in the aMCI group (-0. 75 ± 0. 77 and -1. 13 ± 0. 56) and the dMCI group (-1. 07 ± 0. 79 and-1. 00±0. 52) were significantly lower than those of the CN group (0. 46±0. 73 and 0. 60±0. 61),and the difference was statistically significant (P<0. 01). Depression and anxiety in the LLD group (1. 00±0. 53 and 0. 93±0. 98) and the dMCI group (0. 86±0. 80 and 0. 78±0. 82) were significantly higher than those of the CN group (-0. 92±0. 25 and -0. 74±0. 22),and the difference was statistically significant (P<0. 01). Brain network analysis showed that separated neural circuits were implicated in the depression and cognitive im-pairment. Importantly,interactive effects of depression and MCI on the AFCN were also identified,especially in the bilateral somatomotor area,inferior parietal cortex/precuneus,posterior cingulate cortex,right medial prefrontal cortex/dorsolateral prefrontal cortex and hippocampus. Behavioral significance of AFCN also re-vealed the distinctive neural circuits involved in the depression severity and memory deficits,respectively. Conjunction analysis further identified the overlapped neural circuits associated with depression and memory deficits were primarily in the left DLPFC,insula,hippocampus,right inferior prefrontal cortex and dorsomedi-al prefrontal cortex. Conclusions Depression and cognitive impairment synergistically facilitate functional decoupling of AFCN and thus compromise the integrity of amygdala networks. Distinct depression-related or MCI-related neural constructs represent the characteristics of clinical phenotype of depression or MCI alone, while overlapped circuits probably reveal the neural basis of comorbidity of LLD and MCI.