Objective:To investigate the effect of equol on the proliferation of colom cancer cells and to explore the mechanisms.Methods: Colon cancer cells (DLD1,HCT15,COLO205,LOVO,SW480) were incubated, the cell proliferation was identified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay.Reverse transcription PCR and Western blot were used to measure the mRNA and the protein expression of estrogen receptor and nuclear factor (erythroid-derived 2)-like 2 (Nrf2)in the colon cancer cells, respectively.Moreover, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay was used to investigate the effect of estrogen receptor(ER) inhibitor,ERα agonist, and estrogen receptor ERβagonist on the cell proliferation.Results: ERα was faintly expressed in the DLD-1 and HCT-15 cells.However, ERβ expression in DLD1, HCT15, COLO205, LOVO, and SW480 colon cancer cells.Different concentrations of equol (0, 0.5, 1, 5, 10 μmol/L) significantly inhibited the growth of HCT-15 cell with the expression of ERα and ERβ.More-over, different concentrations of equol (0, 0.5, 1, 5, 10 μmol/L) significantly inhibited the growth of LOVO, and SW480 cells with the ERβ expression in a dose-dependent manner as demonstrated with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell proliferation assay.mRNA expressions of ERα and ERβ in HCT-15 were stimulated significantly.Western blotting proved that the protein expressions of ERα and ERβ increased with the increasing of equol dose.Moreover we found significant difference of Nrf2 protein expression in HCT-15 cell stimulated by different concentrationss of equol.After the similation of estrogen receptor inhibitor, ERα agonist, or ERβ agonist, we found that only dif-ferent concentrations of ERβ agonist(0, 1, 10, 100, 1 000, 10 000 nmol/L) significantly inhibited the growth of HCT-15, LOVO, and SW480 in adose-dependent manner.Estrogen receptor inhibitor and ERα agonistdid not present significant effect on the cell proliferation of HCT-15, LOVO, and SW480.Conclusion: Equol inhibited the colon cancer cell proliferation by its estrogenic activities and antioxidant activities.

Objective To investigate the effects of recombinant human osteoprotegerin(rhOPG) on RANKL ,OPG protein expression in alveolar bone tissue of rats with periodontitis to provide the experimental evidence for the application of rhOPG in pe‐riodontitis treatment .Methods Totally 22 Wistar rats were enrolled .The random number table was adopted to select two healthy rats as the healthy group .The rest 20 rats were selected as the experimental group for establishing the rat models of periodontitis , and then subdivided into the experimental control group (n=10) and rhOPG group (n=10) .Rats in the rhOPG group were locally injected by rhOPG 10 mg/kg at periodontal pocket gap of maxillary second molar ,while those in the experimental control group were injected by sterile water for injection at the same site and some volume .The streptavidin‐perosidase(SP) method was em‐ployed to detect the expression of RANKL ,OPG protein in alveolar bone tissue .Results Compared with the healthy group ,the ex‐pression levels of OPG in alveolar bone tissue of rats in the experimental group were lower with statistically significant difference (P<0 .05) ,while the difference of RANKL expression levels between the two groups showed no statistical significance(P>0 .05) . Compared with the experimental control group ,the expression level of OPG protein in alveolar bone tissue of rats in the rhOPG group was significantly up‐regulated ,while that of RANKL protein was significantly down‐regulated(P<0 .05) .The OPG expres‐sion level after treatment in the rhOPG group was markedly enhanced ,while the RANKL expression level was reduced compared with before treatment ,the difference was statistically significant (P<0 .05) .Conclusion rhOPG may regulates the expression of RANKL and OPG in alveolar bone tissue of rats with periodontitis .

Objective To study the expression of WWOX and C-JUN in keloid and to approach their role and mechanism in the pathogenesis of keloid.Methods Immunohistochemical SP methods were used with computer pathological image analysis.Western blot and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of WWOX and C-JUN in keloid and normal skin with statistical analysis.Results In keloid,the expression of WWOX protein was located in the cytoplasm of fibroblasts,and the expression of WWOX protein and its mRNA decreased,with significantly statistical difference (P＜0.05) compared to normal skin in the control group; the expression of C-JUN protein was located in the cell nucleus and cytoplasm of fibroblasts,with increased expression of C-JUN protein and its mRNA,with significantly statistical difference (P＜0.05) in comparison to normal skin in the control group.The expression of both was negative correlation (r=-0.626,P＜0.01).Conclusions Both WWOX with low expression and C-JUN with high expression are keloid-related genes,having significantly negative correlation between them,which may be one of the mechanisms for the keloid formation.It indicates that the WWOX protein may be an inhibitory factor to the expression of C-JUN protein,and the genes may play a major role in the pathogenesis of keloiod through fibroblasts.

AIM: To study the suppressive effect of glycogen synthase kinase-3β( GSK-3β) knockdown by RNA interference on the formation of keloid .METHODS:Human keloid fibroblasts ( KFB) in vitro were transfected with 3 pairs of specific GSK-3βsmall interfering RNA (siRNA).The best siRNA to inhibit the GSK-3βexpression in human KFB was screen by RT-PCR and Western blot .The expression of GSK-3βand related proteins at mRNA and protein levels in the KFB was determined by RT-PCR and Western blot .RESULTS: The GSK-3βsiRNA1434 remarkably inhibited the expression of GSK-3βat mRNA and proteins levels in the human KFB .After transfection with GSK-3βsiRNA, the protein levels of β-catenin, p-GSK-3β, Wnt2 and cyclin D1 were all decreased.KFB growth became slow.With the extension of time, the inhibition of cell growth increased , and the cell doubling time was significantly delayed .CONCLUSION:siRNA targeting GSK-3βefficiently knocks down the expression of GSK-3βin the human KFB, and inhibits the activation of Wnt signaling pathway , thus inhibiting the growth of keloid .GSK-3βmay be a potential therapeutic target for keloid .

Objective To study the expression of eukaryotic translation initiation factor 4E(eIF4E)in the pathological scars and its probable role in the pathogenesis of pathological scars.Methods Immunohistochemiscal technique was performed to detect the expression and distribution of eIF4E protein in hypertrophic scars(14 cases),keloids(25 cases),mature scars(20 cases)and normal skins(20 cases).Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the eIF4E mRNA level in hypertrophic scars(7 cases),keloids(8 cases),mature scars(8 cases)and normal skins(8 cases).Results Thepositive rate of eIF4E protein expression was remarkably significant difference between normal scars and pathological scars(P＜0.05).The level of eIF4E mRNA in pathological scars 1.73±0.31was higher than that in control group 0.99±0.28.There was significant difference between two groups (P＜O.05).Conclusions The expression of eIF4E is increased in pathological scar.eIF4 E expression is closely associated with the development of pathological scar.Therefore,eIF4E overexpression may play an important role in the proliferation of fibroblasts and in the pathogenesis of pathological scar.

[Objective]To explore the expression of transcription factor KLF16 in nonalcoholic fatty liver disease(NAFLD)and its effect on lipid metabolism.[Methods]An animal model of NAFLD was constructed in mice induced by a high-fat diet.The mice were divided into normal diet group(ND)and high fat diet group(HFD).NAFLD cell model was constructed by primary mouse liver cells induced by oleic acid.The cells were divided into control group(Control group)and oleic acid induction group(OA group).Real-time fluorescence quantitative PCR(RT-qPCR)and Western blot were used to detect KLF16 expression in NAFLD animal and cell models.In vitro and in vivo models of KLF16 knockdown were constructed by injection of adeno-associated virus(AAV)into mouse tail veins and transient transfection of cell siRNA.Hematoxylin-eosin staining(HE)and other methods were used to detect changes in lipid deposition in NAFLD models be-fore and after KLF16 knockout.RT-qPCR was used to detect the expression of key genes of lipid metabolism in both cellu-lar and animal NAFLD models before and after KLF16 knockdown.Western blot assay was used to detect the expression of endoplasmic reticulum stress protein in NAFLD model before and after KLF16 knockdown.[Results]The expression level of KLF16 was up-regulated in HFD group and OA group,and lipid deposition was increased in OA group after KLF16 was depressed.There was no change in TC level in hepatocytes between groups(P>0.05),and TG level was increased in differ-ent degrees(P<0.05,P<0.001).At the same time,the change of KLF16 expression also caused the change of ER stress protein expression in OA group.[Conclusion]The transcription factor KLF16 may alleviate lipid deposition in nonalcoholic fatty liver disease by endoplasmic reticulum stress.

Objective:To investigate the cognitive functions of temporal lobe epilepsy (TLE) patients with bilateral asynchronous interictal discharges.Methods:A total of 162 TLE patients who were treated at Zhongshan Hospital, Fudan University, from June 2021 to December 2023 were collected. According to the interictal scalp electroencephalogram, TLE patients were classified to the TLE with bilateral temporal asynchronous interictal epileptiform discharges ( n=51) and TLE with unilateral temporal epileptiform discharges ( n=111). Unilateral TLE patients were divided into TLE with right ( n=48) and left ( n=63) temporal epileptiform discharges. The Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), Digital Span (DS), Verbal Fluency Test (VFT), the third part of Color Word Test (CWT-C), Trail Making Test-B (TMT-B), Symbol Digit Modalities Test (SDMT), Auditory Verbal Learning Test (AVLT), Rey-Osterrieth Complex Figure (ROCF), and Similarity Comprehension Test were performed for the participants. The differences of cognitive functions between patients with bilateral and unilateral temporal discharges were compared. Univariate and multivariate Logistic regression models were used to analyze the risk factors for patients with bilateral temporal asynchronous discharges. Spearman analysis was used to explore the correlation between cognitive function and clinical indicators. Results:In the group of TLE patients with bilateral asynchronous discharges compared to those with unilateral discharges, the completion time of CWT-C [67 (55, 103) s vs 59 (50, 71) s, Z=-2.904, P=0.004], TMT-B [159 (108, 219) s vs 129 (95, 180) s, Z=-2.361, P=0.018] was longer. Additionally, TLE patients with bilateral asynchronous discharges got lower scores of MMSE [28 (26, 29) vs 29 (28, 30), Z=3.098, P=0.002], MoCA [23 (19, 28) vs 27 (23, 28), Z=3.175, P=0.001], AVLT1+2+3 [16.843±6.482 vs 19.162±5.526, t=-2.347, P=0.020], AVLT6 [6 (3, 10) vs 8 (5, 10), Z=3.275, P=0.001], ROCF2 [15 (8, 22) vs 20 (12, 25), Z=2.870, P=0.004], ROCF3 [14 (8, 22) vs 20 (11, 25), Z=2.634, P=0.008], and Similarity Test [13 (8, 18) vs 16 (12, 20), Z=2.387, P=0.017] as well as lower VFT-vegetable and fruit count [15 (13, 19) vs 18 (15, 21), Z=2.402, P=0.016] and SDMT completion count [41 (30, 53) vs 51 (40, 60), Z=3.089, P=0.002]. The multivariate Logistic regression analysis showed that the decrease in AVLT6 scores ( OR=1.546, 95% CI 1.150-2.078, P=0.004) and longer TMT-B time ( OR=1.013, 95% CI 1.001-1.025, P=0.035) were independent risk factors for TLE patients with bilateral asynchronous discharges. Conclusions:Compared to the patients with TLE characterized by unilateral temporal lobe discharges, those with asynchronous discharges in bilateral temporal lobes show statistically significant declines in all domains of cognitive functions, including executive function, memory, and language abilities. Decreased cue recall ability in language memory and prolonged trail-making test in executive function are independent cognitive impairment risk factors for bilateral temporal asynchronous discharges.

Objective: To assess the effectiveness of online and offline myopia prevention and control health education interventions using wearable behavior monitoring tools for non myopic elementary school students,so as to provide evidence based medical support for public health practices. Methods: From May to June in 2021, two schools were selected within the same county in Jiangsu Province. School 1 conducted online and offline parental health education ( n =111), while school 2 exclusively conducted offline health education activities, representing the traditional intervention group ( n =122). Students from both schools underwent monitoring through wearable behavior tracking tools, with feedback reports provided (eye distance, eye duration, ambient light, and outdoor exposure time). Both schools relied on activities to carry out health education interventions, and organized the distribution of promotional materials and display boards. The intervention group also established WeChat groups to conduct online "Healthy Parents Action" (answering and providing feedback on health knowledge related to myopia prevention and control, myopia prevention and control, science popularization, etc. raised by parents). Evaluation criteria included myopia rates, post dilation refractive error, and axial length, with a tracking period of two years (from 2021 to 2023). Additionally, the study collected refractive parameters from non myopic students who did not participate in wearable tool monitoring in the 12 classes across the two schools. Results: The baseline results indicated that there were no significant differences between the two groups in terms of refractive parameters and wearable tool monitoring results (including screen time, viewing distance, outdoor exposure time, and homework light exposure)( t/Z/χ 2=1.94,1.17,0.58,0.40,0.80,0.69,0.32, P >0.05). After a two-year follow up, in the first and second year, the myopia rate of the online Healthy Parents Action group (11.4%, 29.7%) were lower than that of the traditional group (26.2%, 50.9%), and the degree of refractive change in the intervention group [0.63(0.38,1.19)D] was lower than that of the traditional group [0.91(0.40,1.50)D], and all the differences were statistically significant( χ 2/ Z =4.93，10.37，2.29， P <0.05). However, there were no significant differences ( P >0.05) in axial length changes between the two groups over the twoyear intervention period. Nevertheless, in the second year, the axial length change in the traditional group [0.35(0.20,0.65)mm] was lower than that in the natural observation group [0.55(0.30,0.75)mm], and this difference was statistically significant ( Z =1.92, P <0.05). Conclusions Online and offline myopia prevention and control health education can effectively reduce myopia rates. The intervention mode combining wearable behavior monitoring tools with online health education may have better effects, but further large sample and multi center studies are needed to provide additional evidence and confirmation.

Objective:To evaluate the relationship between regional coronary flow reserve (CFR) obtained from cadmium-zinc-telluride SPECT(CZT SPECT) myocardial functional perfusion imaging (MFPI) and invasive fractional flow reserve (FFR) measured during coronary angiography (CAG) and its clinical value in guiding coronary interventions.Methods:Forty-two patients (30 males, 12 females, age (63.3±9.8) years) who completed CZT SPECT MFPI in the First Affiliated Hospital of Nanjing Medical University from June 2022 to September 2022 and underwent CAG within 3 months were included retrospectively. The concordance of CFR and FFR for diagnosing myocardial ischemia (CFR<2.0 and FFR<0.8) was calculated at the vascular level. The diagnostic efficacy of coronary stenosis≥70% for decreased myocardial blood flow (CFR<2.0) was calculated. Kappa test was used to analyze the data. Results:A total of 126 major coronary arteries were identified in 42 patients, of which 30(23.8%) had a CFR<2.0 by CZT SPECT and 33(26.2%) had stenosis≥70% in CAG. A total of 32 coronary vessels were performed with MFPI CFR and FFR measurements, of which 6 were both decreased and 21 were both normal, so the concordance rate was 84.4%(27/32)( Kappa=0.612, P<0.001). Among 33 coronary vessels with stenosis≥70%, 13 were with CFR≥2.0. Among 30 coronary vessels with CFR<2.0, 10 were with stenosis<70%. When using stenosis≥70% to diagnose CFR decreasing, the sensitivity was 66.7%(20/30), specificity was 86.5%(83/96), positive predictive value was 60.6%(20/33), negative predictive value was 89.2%(83/93), and accuracy was 81.7%(103/126). Conclusions:The concordance between CFR and FFR for the diagnosis of myocardial ischemia is good. Nearly 1/3 of the coronary arteries with decreased CFR have stenosis<70%, whereas nearly 40% of the coronary arteries with stenosis≥70% are not result in myocardial ischemia. Regional CFR determined by CZT SPECT may have potentially significant clinical value in the diagnosis of coronary artery disease and decision-making of coronary intervention.

OBJECTIVE To understand the information labeling of children’s medication in the instructions of antitussive and expectorant drugs commonly used in children’s medical institutions， analyze the existing problems， and propose relevant suggestions. METHODS The instructions for 100 antitussive and expectorant drugs commonly used in 20 tertiary children’s hospitals （centers） and 14 maternal and child health hospitals （centers） with regional representativeness in China were collected， and the information labeling of children’s medication in the instructions was investigated and analyzed. RESULTS There were only 7 kinds of antitussive and expectorant drugs for children， and the others were non-specific drugs for children. Among antitussive and expectorant drugs， tablets accounted for 18.00%， injections for 18.00%， and capsules for 4.00%. Among 100 antitussive and expectorant drugs， 72 （72.00%） labeled the usage and dosage for children， 63 （63.00%） labeled the medication items for children， 59 （59.00%） indicated the information of children in the precautions， and pharmacokinetic parameters for children were absent. Compared with imported antitussive and expectorant drugs or the drugs manufactured by joint ventures， there were many missing labeling of medication information for children with Chinese drugs. Among 63 kinds of drug instructions labeled with medication items for children， various instructions information had little guiding significance. CONCLUSIONS There is a lack of labeling in the instructions of antitussive and expectorant drugs for children， and the proportion of special drugs for children remains low. Pediatric drug information in the instructions has little guiding significance for pediatric medication. Relevant departments should further promote the completeness of pediatric medication information in the instructions of antitussive and expectorant drugs to ensure the rational use of children’s medication.