AIM:To explore the mechanism of PX-12 induced apoptosis of hepatocellular carcinoma cells.METHODS:Human hepatoma cell line Huh7 was se-lected as the main research object.After the cells were treated with thioredoxin inhibitor PX-12,the cell viability was detected by CCK8 method,the cell mi-gration ability was detected by cell scratch test,the cell proliferation ability was detected by cell prolif-eration kit,the levels of intracellular reactive oxygen species and apoptosis were detected by flow cy-tometry,and the expression of apoptosis-related proteins were detected by Western blot.RESULTS:Compared with the control group,the cell viability,migration ability and proliferation ability of PX-12 treatment group were significantly decreased(P＜0.05),and the level of intracellular reactive oxygen species was increased(P＜0.05)in a concentration-dependent manner.Apoptosis inhibitor Z-VAD-FMK and antioxidant NAC could restore the cell viability,and NAC could reduce the accumulation of intracel-lular reactive oxygen species induced by PX-12 and restore the apoptosis induced by PX-12(P＜0.05).CONCLUSION:PX-12 induces apoptosis of hepato-cellular carcinoma cells through oxidative stress.

Digital guided therapy (DGT) has been advocated as a contemporary computer-aided technique for treating endodontic diseases in recent decades. The concept of DGT for endodontic diseases is categorized into static guided endodontics (SGE), necessitating a meticulously designed template, and dynamic guided endodontics (DGE), which utilizes an optical triangulation tracking system. Based on cone-beam computed tomography (CBCT) images superimposed with or without oral scan (OS) data, a virtual template is crafted through software and subsequently translated into a 3-dimensional (3D) printing for SGE, while the system guides the drilling path with a real-time navigation in DGE. DGT was reported to resolve a series of challenging endodontic cases, including teeth with pulp obliteration, teeth with anatomical abnormalities, teeth requiring retreatment, posterior teeth needing endodontic microsurgery, and tooth autotransplantation. Case reports and basic researches all demonstrate that DGT stand as a precise, time-saving, and minimally invasive approach in contrast to conventional freehand method. This expert consensus mainly introduces the case selection, general workflow, evaluation, and impact factor of DGT, which could provide an alternative working strategy in endodontic treatment.
Humans ; Consensus ; Endodontics/methods* ; Tooth ; Printing, Three-Dimensional ; Dental Care ; Cone-Beam Computed Tomography ; Root Canal Therapy

Chaigui granules(CG)are a compound composed of six herbal medicines with significant antidepressant effects.However,the antidepressant mechanism of CG remains unclear.In the present study,we attempted to elucidate the antidepressant mechanism of CG by regulating purine metabolism and purinergic signaling.First,the regulatory effect of CG on purine metabolites in the prefrontal cortex(PFC)of chronic unpredictable mild stress(CUMS)rats was analyzed by ultra high-performance liquid chro-matography tandem mass spectrometry(UHPLC-MS/MS)targeted quantitative analysis.Meanwhile,purinergic receptors(P2X7 receptor(P2X7R),A1 receptor(A1R)and A2A receptor(A2AR))and signaling pathways(nod-like receptor protein 3(NLRP3)inflammasome pathway and cyclic adenosine mono-phosphate(cAMP)-protein kinase A(PKA)pathway)associated with purine metabolism were analyzed by western blotting and enzyme-linked immunosorbent assay(ELISA).Besides,antidepressant mecha-nism of CG by modulating purine metabolites to activate purinergic receptors and related signaling pathways was dissected by exogenous supplementation of purine metabolites and antagonism of puri-nergic receptors in vitro.An in vivo study showed that the decrease in xanthine and the increase in four purine nucleosides were closely related to the antidepressant effects of CG.Additionally,purinergic re-ceptors(P2X7R,A1R and A2AR)and related signaling pathways(NLRP3 inflammasome pathway and cAMP-PKA pathway)were also significantly regulated by CG.The results of exogenous supplementation of purine metabolites and antagonism of purinergic receptors showed that excessive accumulation of xanthine led to activation of the P2X7R-NLRP3 inflammasome pathway,and the reduction of adenosine and inosine inhibited the A1R-cAMP-PKA pathway,which was significantly ameliorated by CG.Overall,CG could promote neuroprotection and ultimately play an antidepressant role by inhibiting the xanthine-P2X7R-NLRP3 inflammasome pathway and activating the adenosine/inosine-A1R-cAMP-PKA pathway.

Radix Bupleuri(RB)is commonly used to treat depression,but it can also lead to hepatotoxicity after long-term use.In many anti-depression prescriptions,RB is often used in combination with Radix Paeoniae Alba(RPA)as an herb pair.However,whether RPA can alleviate RB-induced hepatotoxicity remain unclear.In this work,the results confirmed that RB had a dose-dependent antidepressant effect,but the optimal antidepressant dose caused hepatotoxicity.Notably,RPA effectively reversed RB-induced hepatotoxicity.Afterward,the mechanism of RB-induced hepatotoxicity was confirmed.The results showed that saiko-saponin A and saikosaponin D could inhibit GSH synthase(GSS)activity in the liver,and further cause liver injury through oxidative stress and nuclear factor kappa B(NF-KB)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway.Furthermore,the mechanisms by which RPA attenuates RB-induced hepatotoxicity were investigated.The results demonstrated that RPA increased the abundance of intestinal bacteria with glycosidase activity,thereby promoting the conversion of saikosaponins to sai-kogenins in vivo.Different from saikosaponin A and saikosaponin D,which are directly combined with GSS as an inhibitor,their deglycosylation conversion products saikogenin F and saikogenin G exhibited no GSS binding activity.Based on this,RPA can alleviate the inhibitory effect of saikosaponins on GSS activity to reshape the liver redox balance and further reverse the RB-induced liver inflammatory response by the NF-κB/NLRP3 pathway.In conclusion,the present study suggests that promoting the conversion of saikosa-ponins by modulating gut microbiota to attenuate the inhibition of GSS is the potential mechanism by which RPA prevents RB-induced hepatotoxicity.

Objective:To establish a standard colorectal neoplasm tissue biobank with complete clinical information to provide high quality samples for fundamental and clinical research of colorectal neoplasm.Methods:Based on Affiliated Jinhua Hospital, Zhejiang University School of Medicine, to conduct structural design of colorectal neoplasm tissues, normal tissues and related information. Establish standard operating procedures from the collection and storage of tissue samples, standardize the entry of basic information, medical history, pathology and other relevant clinical information of the patients, and conduct random quality inspections on the pathological morphology and molecular level on a regular basis.Results:A tissue biobank of colorectal neoplasm was successfully constructed. During the establishment and improvement of this tissue biobank, standardized quality control was implemented during the whole-process including sample collection, warehousing, storage and delivery. According to the random sampling quality inspection, the RNA preservation effect was good, the rates of neoplasms in cancer tissue was >80%, and the clinical data of samples were complete.Conclusions:The preliminary construction of colorectal neoplasm tissue biobank not only improves the utilization value of tissue samples, but also provides a guarantee for realizing the bidirectional transformation of fundamental research and clinical application.

Objective To evaluate the effect of amphotericin B on the production of tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) and activation of p38 mitogen-activated protein kinases (p38MAPK) in a human acute monocytic leukemia cell line (THP-1).Methods Cultured THP-1 cells were divided into several groups:blank control group receiving no treatment,amphotericin B groups treated with 2,4 and 8 mg/L amphotericin B separately,positive control group treated with 100 μg/L β-glucosan or 100 mg/L lipopolysaccharide.Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of TNF-α and IL-8 after the THP-1 cells were treated with different stimuli for some durations.Enzyme-linked immunosorbent assay (ELISA) was conducted to detect the level of TNF-α in the culture supernatant of THP-1 cells after 24-hour treatment with 8 mg/L amphotericin B,and Western blot analysis to measure the levels of p38MAPK and phosphorylated p38MAPK after 30-minute treatment with 8 mg/L amphotericin B.Results After 6-hour treatment with 2,4 and 8 mg/L amphotericin B separately,the mRNA levels of TNF-α in THP-1 cells (7.55 ± 1.17,19.47 ± 2.91,57.22 ± 0.65) and IL-8 (2.98 ± 0.04,5.22 ± 1.35,11.82 ± 1.66) were all significantly higher than those in the blank control group (TNF-α:1.00 ± 0.07,P ＜ 0.01,0.001,0.001 respectively;IL-8:1.01 ± 0.23,P ＜ 0.01,0.001,0.001 respectively).After the treatment with 8 mg/L amphotericin B for 1,3,6 hours,the mRNA levels of TNF-α (8.61 ± 0.30,10.75 ± 0.08,56.98 ± 2.43) and IL-8 (2.63 ± 0.28,5.35 ± 0.98,11.73 ± 1.18) in THP-1 cells were all significantly higher than those in the blank control group (TNF-α:1.18 ± 0.17,P ＜ 0.05,0.01,0.001;IL-8:1.23 ± 0.11,P ＜ 0.05,0.01,0.001).After 24-hour treatment with 8 mg/L amphotericin B,the level of TNF-α in the culture supernatant of THP-1 cells was significantly higher than that in the blank control group (4 039.06 ± 223.87 ng/L vs.96.31 ± 0.26 ng/L,P ＜ 0.001).Conclusion Amphotericin B can promote the p38MAPK phosphorylation and increase the levels of TNF-α and IL-8 in human THP-1 cells in vitro,suggesting its immunomodulatory effects.

Whether the drug is successfuly transported to the focus area is the key to the treatment of the rhinitis disease. The efficiency of drug delivery is dependent on the design of nasal spray device. A three-dimensional model of nasal cavity was constructed from the head CT image data of a healthy human subject. The deposition of drug particles was simulated numericaly by Computational Fluid Dynamics technique. The main variables of the study were the size and density of drug particles, the injecting speed of drugs etc. In conclusion, with the increase of particle intensity and injection speed, the deposition of particles in the affected area trends to increase after first slow decrease.

AIM:To investigate the expression of microRNA-193b(miR-193b) in the cervical tissues, and fur-ther to explore the effect of silencing miR-193b on diamminedichloroplatinum ( DDP)-treated HeLa cell viability.METH-ODS:The expression levels of miR-193b in different cervical tissues were examined by qPCR.After transfection of miR-193b-inhibitor, the cell migration was determined by Transwell assay, the sensitivity of HeLa cells to DDP was measured by MTT assay, the protein levels of phosphate and tension homology deleted on chromsome ten ( PTEN), protein kinase B (Akt), p-Akt and p-glycoprotein(P-gp) were determined by Western blot.RESULTS:The mRNA level of miR-193b was significantly increased in the cervical cancer tissues compared with normal cervical tissues ( P<0.05) .Knockdown of miR-193b obviously inhibited migration and enhanced sensitivity to DDP of HeLa cells (P<0.05).Additionally, after transfec-tion of miR-193b-inhibitor, the expression of PTEN was increased, whereas the protein levels of p-Akt and P-gp were de-creased (P<0.05).CONCLUSION:miR-193b is highly expressed in the cervical cancer tissues.Inhibition of miR-193b augments the sensitivity to DDP of HeLa cells, at least in part, through PTEN-PI3K/Akt signaling pathway.

Aim To investigate the effects of diterpene ginkgolides meglumine injection (DGMI ) on amino acids and monoamine neurotransmitters in rats with cerebral ischemia/reperfusion injury.Methods In-traluminal suture was applied to establish middle cere-bral artery occlusion (MCAO/R)model with ischemia for 1.5 h and reperfusion for 24 h.After the adminis-tration of DGMI (i.v.),the levels of amino acid and monoamine neurotransmitters in brain tissue were de-tected through HPLC-ECD.Results DGMI down-reg-ulated the concentrations of aspartic acid, glutamic acid,glycine and γ-aminobutyric acid which were in-creased in MCAO/R group.DGMI also reduced the levels of norepinephrine epinephrine,glyoxylic acid, serotonin and 5-HIAA in cortex and hippocampus,and increased adrenaline content compared to the model group.Conclusion DGMI exhibits a protective role in rats with cerebral ischemia /reperfusion injury through regulating amino acids and monoamine neuro-transmitters.

The purpose of this study is to evaluate the anti-aging effects and reveal the underlying mechanism of Scutellaria baicalensis Georgi ethanol extract (SBG) in D-galactose-induced rats. Fifty rats were randomly divided into five groups: vehicle control group, D-galactose group, and D-galactose combined with 50, 100, 200 mg x kg(-1) SBG. A rat aging model was induced by injecting subcutaneously D-galactose (100 mg x kg(-1)) for ten weeks. At the tenth week, the locomotor activity (in open-field test) and the learning and memory abilities (in Morris water maze test) were examined respectively. The urine was collected using metabolic cages and analyzed by high-resolution 1H NMR spectroscopy combined with multivariate statistical analyses. The SBG at doses of 50, 100 and 200 mg x kg(-1) treatments groups could significantly ameliorate aging process in rats' cognitive performance. The 50, 100, 200 mg x kg(-1) SBG regulated citrate, pyruvate, lactate, trimethylamine (TMA), pantothenate, β-hydroxybutyrate in urine favorably toward the control group. These biochemical changes are related to the disturbance in energy metabolism, glycometabolism and microbiome metabolism, which is helpful to further understanding the D-galactose induced aging rats and the therapeutic mechanism of SBG.