ObjectiveTo investigate the effects of Zuoguiwan on osteoclast activation induced by breast cancer and its mechanism. MethodsTo simulate breast cancer-induced osteoclastic bone metastasis， RAW264.7 cells were cultured in conditioned medium containing 50% supernatant of MDA-MB-231 breast cancer cells. The dosages of Zuoguiwan used in the experiment were sera containing 5% and 10% Zuoguiwan. Tartrate-resistant acid phosphatase （TRAP） staining was used to detect osteoclast activation. Enzyme-linked immunosorbent assay （ELISA） was used to measure Cathepsin K secretion from RAW264.7 cells. Real-time quantitative polymerase chain reaction （PCR） was used to detect the mRNA expression levels of osteocalcin （OCN） and bone sialoprotein （BSP）. Immunoprecipitation was employed to detect the interaction between Runt-related transcription factor 2 （Runx2） and core binding factor β subunit （CBF-β）. Western blot was used to assess the protein expression of Runx2， phosphorylated Runx2 （p-Runx2）， extracellular signal-regulated kinases 1/2 （ERK1/2）， p-ERK1/2， p38 mitogen-activated protein kinase （MAPK）， p-p38 MAPK， and CBF-β. ResultsCompared with the blank group， the MDA-MB-231 cell supernatant group showed a significant increase in TRAP-positive cell counts and Cathepsin K secretion. Meanwhile， the expression levels of p-Runx2， Runx2-CBF-β interaction， BSP and OCN mRNA， p-p38 MAPK， and p-ERK1/2 proteins were significantly decreased （P<0.01）. Compared with the MDA-MB-231 cell supernatant group， Zuoguiwan-containing sera significantly reduced TRAP-positive cell counts and Cathepsin K secretion （P<0.01）， significantly increased p-Runx2， BSP and OCN mRNA expression， as well as p-p38 MAPK and p-ERK1/2 protein levels， and promoted the interaction between Runx2 and CBF-β （P<0.01）. No significant change in Runx2 expression was observed. Compared to the blank group， the BVD-523 group showed significantly lower expression of p-p38 MAPK and p-ERK1/2 proteins （P<0.01）. Compared with the BVD-523 group， both low and high concentration Zuoguiwan-containing sera groups showed significantly higher p-p38 MAPK expression （P<0.01）， and the high concentration Zuoguiwan group also exhibited a significant increase in p-ERK1/2 expression （P<0.01）， while no statistical difference was found in the low-dose group. ConclusionZuoguiwan inhibits osteoclast activation by inducing phosphorylation of the key transcriptional regulator Runx2 in intra-osteoclast bone formation， and this process is closely associated with the activation of the p38 MAPK/ERK signaling pathway.

Objective:To quantitatively evaluate the accuracy of data obtained from liquid-interference surfaces using an intraoral 3D scanner(IOS)integrated with a compressed airflow system,so as to pro-vide clinical proof of accuracy for the application of the compressed airflow system-based scanning head in improving data quality on liquid-interference surfaces.Methods:The study selected a standard model as the scanning object,adhering to the"YY/T 1818-2022 Dental Science Intraoral Digital Impression Scanner"guidelines,a standard that defined parameters for intraoral scanning.To establish a baseline for accuracy,the ATOS Q 12M scanner,known for its high precision,was used to generate true reference values.These true values served as the benchmark for evaluating the IOS performance.Building on the design of an existing scanner,a new scanning head was developed to integrate with a compressed airflow system.This new design aimed to help the IOS capture high-precision data on sur-faces where liquid-interference,such as saliva,might otherwise degrade scanning accuracy.The tradi-tional scanning method,without airflow assistance,was employed as a control group for comparison.The study included five groups in total,one control group and four experimental groups,to investigate the effects of scanning lens obstruction,airflow presence,liquid media,and the use of the new scan-ning head on scanning process and accuracy.Each group underwent 15 scans,generating ample data for a robust statistical comparison.By evaluating trueness and precision in each group,the study as-sessed the impact of the compressed airflow system on the accuracy of IOS data collected from liquid-interference surfaces.Additionally,we selected Elite and Primescan scanners as references for numeri-cal accuracy values.Results:The scanning accuracy on liquid-interference surfaces was significantly reduced in terms of both trueness and precision[Trueness:18.5(6.5)vs.38.0(6.7),P＜0.05;Preci-sion:19.1(8.5)vs.31.7(15.0),P＜0.05].The use of the new scanning head assisted by the com-pressed airflow system significantly improved the scanning accuracy[Trueness:22.3(7.6)vs.38.0(6.7),P＜0.05;Precision:25.8(9.6)vs.31.7(15.0),P＜0.05].Conclusion:The scanning head based on the compressed airflow system can assist in improving the accuracy of data obtained from liquid-inter-ference surfaces by the IOS.

Cetuximab is a monoclonal antibody (mAb) targeting the epidermal growth factor receptor (EGFR) for the treatment of metastat-ic colorectal cancer (mCRC). However,most patients treated with cetuximab experience disease progression due to the development of sec-ondary drug resistance,presenting a significant challenge in managing cetuximab therapy. Existing studies have found that cetuximab resist-ance mechanisms are not only linked to the RAS/RAF/PIK3CA genetic mutations but also closely associated with the abnormal activation of PI3K/AKT/mTOR,Wnt/β-catenin,c-MET/HGF,and RAS-MAPK signaling pathways. Additionally,HER2 and MET amplification,microsatellite instability,changes in tumor metabolism,and alterations in the tumor microenvironment may also contribute to cetuximab resistance in pa-tients. This review focuses on the potential molecular mechanisms of cetuximab resistance in the treatment of mCRC,and provides new ideas for overcoming cetuximab resistance in clinic.

Cetuximab is a monoclonal antibody (mAb) targeting the epidermal growth factor receptor (EGFR) for the treatment of metastat-ic colorectal cancer (mCRC). However,most patients treated with cetuximab experience disease progression due to the development of sec-ondary drug resistance,presenting a significant challenge in managing cetuximab therapy. Existing studies have found that cetuximab resist-ance mechanisms are not only linked to the RAS/RAF/PIK3CA genetic mutations but also closely associated with the abnormal activation of PI3K/AKT/mTOR,Wnt/β-catenin,c-MET/HGF,and RAS-MAPK signaling pathways. Additionally,HER2 and MET amplification,microsatellite instability,changes in tumor metabolism,and alterations in the tumor microenvironment may also contribute to cetuximab resistance in pa-tients. This review focuses on the potential molecular mechanisms of cetuximab resistance in the treatment of mCRC,and provides new ideas for overcoming cetuximab resistance in clinic.

Objective To search for novel potent 3-ester derivatives of arenobufagin and test their antitumor activities in vitro. Methods Target compounds were synthesized by esterification of arenobufagin with acids. CellTiter method was used to assay the in vitro antitumor activities. Results 3-Ester derivatives exhibited excellent antitumor activities against all the cancer cells. Conclusion Among the 3-ester derivatives, compound 2a had the best activities with the IC₅₀ of 4.0−91.7 nmol/L and appeared to be a valuable candidate for further study.

Objective:To report 1 case of mushroom poisoning complicated with multiple organ dysfunction syndrome treated by liver transplantation, so as to explore surgical effectiveness and prognosis of liver transplantation on children mushroom poisoning.Methods:Retrospectively analyzed a child with mushroom poisoning who was admitted to the First Affiliated Hospital of Zhengzhou University in June 2017.Conventional treatment was not ideal, the child was conducted orthotopic liver transplantation.The effectiveness was evaluated and the feasibility of liver transplantation in treating children mushroom poisoning with literatures was reviewed.Results:A male 2-year old child obtained, preo-perative Child-pugh class for C. This child recovered well postoperatively, the jaundice disappeared, and liver enzyme and blood coagulation function improved significantly within 1 week after liver transplantation.However, the head magnetic resonance imaging presented brain atrophy and ventricular expansion, with intelligence and athletic ability being significantly behind his peers through postoperative regular follow-up for 2 years.Conclusion:Fulminant hepatic failure that was caused by mushroom poisoning is an indication for liver transplantation, and orthotopic liver transplantation is an effective method for the treatment of mushroom poisoning, which can effectively improve liver function, metabolic disorders and blood coagulation function for children with mushroom poisoning, but nervous system damage that was caused by mushroom poisoning is irreversible.Although liver transplantation technology successfully cured liver failure, it still left severe neurological sequelae.

Objective:To evaluate the role of hypoxia-inducible factor-1α (HIF-1α)/Bcl-2/E1B-19kDa interacting protein 3 (BNIP3) signaling pathway in sevoflurane-induced attenuation of myocardial ischemia-reperfusion (I/R) injury in rats and the relationship with autophagy.Methods:Ninety healthy adult male Sprague-Dawley rats, weighing 300-350 g, were randomly divided into 5 groups ( n=18 each): sham operation group (Sham group), I/R group, sevoflurane group (SEV group), HIF-1a inhibitor 2ME2 group (2ME2 group), and 2ME2+ sevoflurane group (MSP group). Myocardial I/R injury model was established by ligating the left anterior descending branch of coronary artery for 40 min followed by 120-min reperfusion in anesthetized rats.In SEV group, 2.4% sevoflurane was inhaled for 15 min starting from the beginning of reperfusion.In 2ME2 group and MSP group, 2ME2 (15 mg/kg) was intraperitoneally injected at 1 h before ligation of the left anterior descending branch of coronary artery, and the other treatments were similar to those previously described in group I/R or in group SEV.Animals were sacrificed at 120 min of reperfusion, and the left ventricular myocardium was taken for determination of the expression of HIF-1α and BNIP3 (by Western blot), activity of ROS (by DHE), and myocardial infarct size (by TTC)and for observation of autophagosome (with an electron microscope). Results:Compared with Sham group, the activity of ROS, the number of autophagosome and myocardial infarct size were significantly increased in the other four groups, the expression of HIF-1α and BNIP3 was up-regulated in I/R group and SEV group ( P<0.05). Compared with I/R group, the activity of ROS, the number of autophagosome and myocardial infarct size were significantly decreased in the other four groups, the expression of HIF-1α and BNIP3 was up-regulated in SEV group, and no significant difference was found in the activity of ROS, the number of autophagosome or myocardial infarct size ( P>0.05), and the expression of HIF-1α and BNIP3 was down-regulated in 2ME2 and MSP groups ( P<0.05). Compared with SEV group, the activity of ROS, the number of autophagosome and myocardial infarct size were significantly increased, and the expression of HIF-1α and BNIP3 was down-regulated in 2ME2 and MSP groups ( P<0.05). Conclusion:HIF-1α/BNIP3 signaling pathway is involved in sevoflurane-induced attenuation of myocardial I/R injury, which is related to inhibiting autophagy in rats.

Objective To study the expressions and clinical significances of activating transcription factor 3 (ATF3) and Runt-related transcription factor 2 (Runx2) in breast carcinoma tissues.Methods The expressions of ATF3 and Runx2 were detected in 105 cases of primary breast invasive ductal carcinoma (IDC) and their matched para-tumor breast tissues by immunohistochemistry.The relationships between the two transcription factors and the clinical pathological features of IDC patients were analyzed.Results The positive expression rates of ATF3 and Runx2 in IDC group were 80.95％ (85/105) and 69.52％ (73/105) respectively,much higher than those in para-tumor breast tissues 11.43 ％ (12/105) and 8.57％ (9/105),with statistically significant differences (x2 =102.097,P =0.000;x2 =11.595,P =0.001).ATF3 and Runx2 expressions showed significant relationships with histological grade (x2 =14.623,P =0.001;x2 =24.891,P =0.000),lymph node metastasis (x2 =7.059,P =0.008;x2 =6.358,P =0.012) and pTNM stage of IDC (x2 =5.807,P =0.016;x2 =4.902,P =0.027),while both were not correlated with patients' age (x2 =0.274,P =0.601;x2 =1.554,P =0.213) and tumor size (x2 =2.476,P =0.290;x2 =5.261,P =0.072).There was a significant positive relationship between ATF3 and Runx2 in breast carcinoma (C =0.498,P =0.000).Conclusion The over-expressions of ATF3 and Runx2 may participate in the tumorigenesis,invasion,metastasis and clinical stage of breast carcinoma,which suggests that they may be key factors to evaluate malignant degree and prognosis of breast carcinoma.

BACKGROUNDHerpes simplex keratitis (HSK) caused by herpes simplex virus 1 (HSV-1), which has high recurrent rate and incidence of severe vision loss, is the leading cause of infectious blindness in the world. The aim was to explore the clinical efficacy of oral ganciclovir (GCV) in the prevention of recurrent HSK.

METHODSA multicenter, prospective, randomized, single-blind, and controlled clinical trial was conducted from April 2010 to June 2013. One hundred seventy-three patients (173 eyes involved) who were diagnosed as recurrent HSK definitely, including stromal keratitis and corneal endotheliitis, were divided into three groups randomly: negative control (placebo) group was topically administered with 0.15% GCV ophthalmic gel, 4 times per day and 0.1% fluorometholone eye drops, 3 times per day until resolution of HSK; positive control acyclovir (ACV) group was topically adopted the same ophthalmic gel and eye drops and additionally received oral ACV 400 mg 5 times a day for 10 weeks and followed by 400 mg 2 times per day for 6 months; test GCV group was topically adopted the same treatment as negative control group and additionally received oral GCV 1000 mg 3 times per day for 8 weeks. The symptoms and signs were evaluated before and after the therapy 1 st week, 2 nd week and then followed up every 2 weeks until recovery. Furthermore, we followed up recurrence of HSK for every 3 months after recovery and then assessed the cure time, recurrent rate and adverse reactions.

RESULTSOne hundred and seventy-three patients were followed up 7-48 months (mean 32.1 ± 12.3 months), but 34 patients were failed to follow-up. The cure time was 12.1 ± 4.3, 11.9 ± 4.0 weeks in negative control (placebo) group and positive control ACV group respectively (P = 0.991), which was longer than that in test GCV group (8.6 ± 2.8 weeks) and there was a significant difference between test GCV group and negative control (placebo) group or positive control ACV group (P = 0.000). Furthermore, the recurrent rate was higher in negative control (placebo) group (47.3%) than that in positive control group ACV (26.7%) and test GCV group (17.2%), and there was a great significant difference among the three groups (P = 0.007), but there was no significant difference between positive control ACV group and test GCV group (P = 0.358). In addition, there was no obvious adverse reaction expect neutropenia (only one patient in test GCV group).

CONCLUSIONShort-term oral GCV could cure recurrent HSK and endotheliitis, shorten the course, reduce recurrent rate of HSK and have confirmed safety.

Adult ; Aged ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Ganciclovir ; administration & dosage ; therapeutic use ; Humans ; Keratitis, Herpetic ; drug therapy ; Male ; Middle Aged ; Single-Blind Method ; Treatment Outcome

OBJECTIVETo test expression of carbonic anhydrase II (Ca II) mRNA in osteoclasts which were applied with fluid shear stress.

METHODSThe bone marrow cells of Sprague-Dawley rats were cultured with the presence of 1,25-(OH)2D3 and dexamethasone. The osteoclast-like cells were identified by tartrate-resistant acid phosphatase(TRAP) staining and scanning electron microscope (SEM) observation, then purified with trypsin/ethylenediamine tetraacetic acid (EDTA). Different values and lasting time of steady fluid shear stress were exerted on the osteoclasts with parallel plate flow system. The Ca II expression of osteoclasts were detected by real time reverse transcription-polymerase chain reaction(RT-PCR) and nested polymerase chain reaction(PCR).

RESULTSThe levels of Ca II mRNA were down-regulated correspondingly with the increase of stress and time (P < 0.05).

CONCLUSIONIt's indicated that steady fluid shear stress within a certain range may down-regulate the expression of Ca II in osteoclasts.

Animals ; Bone Marrow Cells ; Carbonic Anhydrase II ; Cells, Cultured ; Osteoclasts ; Polymerase Chain Reaction ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Stress, Mechanical