Objective:To investigate the proliferation of rat glomerular mesangial cells (GMC) influenced by different ethanol extracts of Tripterygium wilfordii HooK F.(TWHF).Methods:An HPLC method was used to establish the fingerprints of 5 different ethanol extracts of TWHF,and GMC was chosen to study the effects of different ethanol extracts of TWHF on cell proliferation.After statistical analysis,the spectrum-activity relationship was analyzed by using partial least squares regression(PLSR).Results:The HPLC fingerprints of the 5 different ethanol extracts of TWHF were established,and 32 characteristic peaks were characterized by the HPLC fingerprints.60%,70% and 95% ethanol extracts and glycosides tablets showed dose-effect relationship,and with the increase of dose,the more significant inhibition of cell proliferation was exhibited.The absorbance values of the 60% ethanol extracts at medium and high doses were lower than those of the other extracts at the same dose.The proliferation inhibition rate of GMC was used as the potency index and analyzed by PLSR,and 20 peaks were potency peaks at high dose(40 μg·L-1),17 ones were potency peaks at medium dose(20 μg·L-1) and 15 ones were potency peaks at low dose(10 μg·L-1).Conclusion:Part of the potency peaks has regular dose-effect relationship with the changes of dose.

Objective:To investigate the mechanism of subjective cognitive decline(SCD)in resting-state by using regional homogeneity(ReHo)and functional connectivity(FC)in SCD patients.Methods:Resting-state functional magnetic resonance imaging(RS-fMRI)was performed in 25 SCD patients and 30 normal controls matched by sex, education and nationality.DPARSFA2.3 and SPM8 software were used to analyze and screen the brain areas with abnormal ReHo values in SCD group, with the posterior cingulated(PCC)/paruneus as seed points for whole-brain FC analysis.Results:Compared with the normal control group, the SCD group showed that ReHo values of right occipital gyrus and left precuneus were increased, and ReHo values of right inferior temporal gyrus, right orbital inferior frontal gyrus and bilateral thalamus were decreased(Voxel level, Alphasim correction, P<0.05). Using PCC/ precuneus as seed voxels, the whole brain functional connectivity analysis showed that the functional connectivity with cerebelum Crus 2 R was increased, and the functional connectivity with right orbital inferior frontal gyrus, left inferior temporal gyrus and temporal pole was reduced(Voxel level, Alphasim correction, all P<0.05). Conclusions:Default mode network may play an important role in the mechanism of SCD, and abnormalities in brain areas may first occur in PCC/precuneus.

Objective:To investigate the characteristics of the nocturnal melatonin secretion concentration and circadian rhythm in patients with wake-up stroke (WUS).Methods:Patients with acute ischemia stroke (AIS) admitted to the Department of Neurology, the Second Affiliated Hospital of Soochow University from October 2019 to August 2022 were enrolled. They were divided into WUS group and non-WUS group. Saliva samples within one week after admission were collected (at 19∶00, 20∶00, 21∶00, 22∶00, and 23∶00) and melatonin concentration was measured. Melatonin secretion curve graph was drawn, dim light melatonin onset (DLMO) was calculated, and circadian rhythms were evaluated. The differences in endogenous circadian rhythms between the WUS group and the non-WUS group were compared. The relevant factors of WUS were determined by multivariate logistic regression analysis. Results:A total of 116 patients with AIS were included, with 79 males (68.1%), aged 59.9±10.3 years; 35 patients (30.2%) were WUS. Univariate analysis showed that there was a statistically significant difference in the infarct site between the WUS group and the non-WUS group ( P=0.019). At 21:00 ( P=0.004) and the average ( P=0.038) nighttime melatonin concentration in the WUS group were significantly lower than those in the non-WUS group, and DLMO showed a significant delay compared to the non-WUS group (21:28:08 vs. 20:57:57; P=0.015). Multivariate logistic regression analysis showed a significant independent correlation between DLMO delay and WUS (odds ratio 1.792, 95% confidence interval 1.123-2.858; P=0.014). Conclusion:Patients with WUS may have endogenous circadian rhythm delay, which is an independent risk factor for WUS.

Objective:To investigate regional homogeneity(ReHo)and whole brain functional connectivity(FC)in patients with mild cognitive impairment(MCI), and to explore the mechanisms of MCI in the resting state.Methods:Resting-state functional magnetic resonance imaging(RS-fMRI)was performed on 24 patients with MCI and 30 age, gender and nationality-matched normal controls.Abnormal brain areas in the MCI group were screened and analyzed by using the DPARSFA2.3 and SPM8 software programs.Whole brain FC analysis was performed with the posterior cingulate cortex(PCC)/precuneus as the seed points.Results:Compared with the control group, MCI subjects displayed higher ReHo values in the frontal-middle-Left, precentral-Left, postcentral-Left, rolandic-opercular-Left, and frontal-inferior-opercular-Left and lower ReHo values in the temporal-superior-right, temporal-middle-right, postcentral-right, and temporal -pole -superior -right(Voxel level, Alphasim correction, P<0.05). Whole brain FC analysis showed greater functional connectivity of PCC/precuneus with fusiform-right, thalamus-right, lingual-right and parahippocampal-right in subjects with MCI, and less functional connectivity of the PCC/precuneus with temporal-middle-Left, angular-Left, temporal-superior-Left and occipital-middle-Left in subjects with MCI(Voxel level, Alphasim correction, P<0.05). Conclusions:Abnormalities of the default mode network may be associated with the onset of MCI, and abnormalities in posterior cingulate/precuneus connectivity may be helpful in finding imaging evidence with high sensitivity to MCI.

Head and neck malignant tumor is one of the most heterogeneous diseases.The multi-disciplinary team(MDT)is an essen-tial component for personal precise diagnosis,treatment and integrated care management of oncologic diseases including head and neck malignant tumor.MDT clinical practice is also an important teaching mode for head and neck malignant tumors,but it is limited by time and space in actual teaching.An internet visualization platform was constructed based on the Internet,hospital HIS/PACS/LIS/EMR system,medical visualization screen,oral endoscope,remote consultation platform and other accessible audio and video terminals,and has been applied in MDT clinical teaching of head and neck malignant tumors,allowing medical students to participate in MDT through a networked visualization platform.Medical students will achieve deep learning for the most heterogeneous malignant tumor.MDT sup-ported by the internet visualization platform provides a new pathway for clinical medical education.

Several putative transcription factor binding sites (TFBSs) exist in the PCV2 rep gene promoter. To explore if porcine circovirus type 2 (PCV2) could regulate the viral replication by using these TFBSs, we conducted electrophoretic mobility shift assay (EMSA), DNA-pull down and liquid chromatography-tandem mass spectrometric (LC-MS/MS) assays. EMSA confirmed the binding activity of the rep gene promoter with nuclear proteins of host cells. DNA-pull down and LC-MS/MS identified the porcine transcription factor AP-2δ (poTFAP2δ) could bind the PCV2 rep gene promoter. Dual-luciferase reporter assay, quantitative real-time PCR, Western blotting and indirect immunofluorescent assay demonstrated that poTFAP2δ could not only promote the activity of the rep gene promoter, but also enhance the transcription/translation activity of the rep/cap gene and the virus titer of PCV2 during the entire life cycle of PCV2 infection. This study revealed the molecular mechanism of PCV2 using host proteins to enhance the viral replication, provided a new perspective for studying the pathogenic mechanism of PCV2 from virus and host interactions, and provided a theoretical basis for developing highly effective PCV2 vaccines.
Animals ; Cell Line ; Chromatography, Liquid ; Circoviridae Infections ; Circovirus ; DNA Helicases ; Diabetes Mellitus, Type 2 ; Promoter Regions, Genetic ; Swine ; Tandem Mass Spectrometry ; Transcription Factor AP-2 ; Virus Replication

Alzheimer's disease （AD） is a common chronic degenerative neurological disease in the elderly and is mainly manifested by the impairment of cognition， memory， and behaviors. At present， the etiology and pathogenesis of this disease have not been fully unraveled， and the research on related drugs for alleviating the disease progression is still in clinical trials. Traditional Chinese medicine （TCM） believes that the elderly have reduced visceral function. Deficiency of vital Qi and turbid phlegm obscuring orifices are the core pathogenesis of AD. Qi deficiency and phlegm obstruction run through the whole pathological process of AD， and the important role of therapeutic principles of supplementing Qi and resolving phlegm is emphasized in treatment. In recent years， the Chinese medicinal compounds effective in supplementing Qi and resolving phlegm represented by Kaixin Powder have been widely used in the clinical and basic research of AD. As reported， in addition to the improvement of the cognitive function of AD， it can also reduce β amyloid （Aβ） deposition， inhibit tau hyperphosphorylation， improve neurotransmitter activity， regulate neuronal synaptic plasticity， resist oxidant stress injury， and inhibit the central inflammatory response and neuronal apoptosis. Therefore， this article reviewed and analyzed the theoretical basis of the treatment of AD by supplementing Qi and resolving phlegm and the mechanism of Chinese medicinal compounds effective in supplementing qi and resolving phlegm against AD to provide theoretical support and a scientific basis for the clinical application of TCM in the prevention and treatment of AD.

Diabetic patients with poor glycemic control are exposed to media containing mold spores, and spores enter the body, which may lead to refractory infections. This article combines case and literature reviews, proposes the diagnosis and treatment method of mold infection, and provides some guidances for subjects who long-term exposure to mold groups such as farmers and immunocompromised people.

OBJECTIVE To explore the improvement mechanism of Modified sanhuang ointment on anal ulcers and swelling model rats. METHODS The anal ulcer model of the rat was induced by using glacial acetic acid； the effects of Modified sanhuang ointment low-dose， medium-dose and high-dose groups （185， 370， 740 mg/kg）， western medicine positive control group （Compound carraghenates cream， 1 g/kg） and TCM positive control group （Mayinglong shexiang zhichuang ointments， 1 g/kg） on body weight， area of anal ulcer， grade of anal ulcer were investigated. The other groups of rats were used to induce rectal swelling models with croton oil； the effects of Modified sanhuang ointment low-dose， medium-dose and high-dose groups （185， 370， 740 mg/kg）， western medicine positive control group （Compound carraghenates cream， 1 g/kg） and TCM positive control group （Mayinglong shexiang zhichuang ointments， 1 g/kg） on the rate of rectal and anal swelling， serum contents of inflammatory factors [interleukin 2 （IL-2）， IL-4， tumor necrosis factor （TNF-α）]， pathological morphology of rectal tissue， the expression of transient receptor potential channel V1 （TRPV1） and substance P in rectal tissue and rectal vascular permeability were investigated. RESULTS In Modified sanhuang ointment， the increase in body weight was enhanced， and the area of anal ulcers， as well as the grade of anal ulcers in rats with anal ulcer models， were reduced to varying degrees； rectal tissue damage in rectal swelling model rats was improved； the rate of rectal and anal swelling， the serum contents of inflammatory factors， the expressions of TRPV1 and substance P in rectal tissue， and rectal vascular permeability were all decreased （P＜0.05 or P＜0.01）. The effect of Modified sanhuang ointment was better than that of western medicine positive control and TCM positive control.Modified sanhuang ointment can improve anal ulcers and swelling in rats by reducing the release of inflammatory factors， inhibiting the expression of TRPV1 and substance P.

Cell membrane affinity chromatography has been widely applied in membrane protein (MP)-targeted drug screening and interaction analysis. However, in current methods, the MP sources are derived from cell lines or recombinant protein expression, which are time-consuming for cell culture or purification, and also difficult to ensure the purity and consistent orientation of MPs in the chromatographic stationary phase. In this study, a novel in situ synthesis membrane protein affinity chromatography (iSMAC) method was developed utilizing cell-free protein expression (CFE) and covalent immobilized affinity chromatography, which achieved efficient in situ synthesis and unidirectional insertion of MPs into liposomes in the stationary phase. The advantages of iSMAC are: 1) There is no need to culture cells or prepare recombinant proteins; 2) Specific and purified MPs with stable and controllable content can be obtained within 2 h; 3) MPs maintain the transmembrane structure and a consistent orientation in the chromatographic stationary phase; 4) The flexible and personalized construction of cDNAs makes it possible to analyze drug binding sites. iSMAC was successfully applied to screen PDGFRβ inhibitors from Salvia miltiorrhiza and Schisandra chinensis. Micro columns prepared by in-situ synthesis maintain satisfactory analysis activity within 72 h. Two new PDGFRβ inhibitors, salvianolic acid B and gomisin D, were screened out with K _D values of 13.44 and 7.39 μmol/L, respectively. In vitro experiments confirmed that the two compounds decreased α-SMA and collagen Ӏ mRNA levels raised by TGF-β in HSC-T6 cells through regulating the phosphorylation of p38, AKT and ERK. In vivo, Sal B could also attenuate CCl₄-induced liver fibrosis by downregulating PDGFRβ downstream related protein levels. The iSMAC method can be applied to other general MPs, and provides a practical approach for the rapid preparation of MP-immobilized or other biological solid-phase materials.