Aim To investigate the synergistic sensiti-zation effect of saikosaponin D(SSD)combined with temozolomide(TMZ)on glioblastoma cells(GBM)and its molecular mechanism.Methods The sensitiv-ity of RG-2,U251 and LN-428 GBM cell lines to SSD and TMZ was analyzed by CCK-8 method combined with HE staining,and the optimal compatible concen-tration was screened.The effect of HE staining com-bined with Hoechst fluorescence staining on the prolif-eration of GBM cell line was detected by clonal forma-tion experiment.The autophagosome formation of GBM cells was observed by monodansylcadaverine(MDC)staining.The expression and distribution of endoplas-mic reticulum stress-related factors and apoptosis and autophagy proteins were detected by Western blot and ICC.Results The sensitivity order of GBM cells to TMZ was RG-2＞U251＞LN-428.The results of com-bined administration showed the synergistic inhibitory effect of SSD combined with TMZ on proliferation of GBM cell lines,which was confirmed by cell cloning formation experiment.Compared with the TMZ group,Hoechst fluorescence staining showed a significant in-crease in the number of nuclear bright staining in the combined administration group.MDC fluorescence staining showed that there were more dense green parti-cles in the cytoplasm of SSD/TMZ plus group than that of TMZ group.Western blot results showed that com-pared with TMZ group,the expression of ER stress markers GRP78,CHOP,p-PERK and ATF6 signifi-cantly increased in SSD/TMZ group(P＜0.05).The expressions of apoptosis proteins caspase-12,caspase-9,caspase-3,cleaved caspase-3,Bax and autophagy proteins LC3 and Beclin-1 significantly increased(P＜0.05),which were verified by ICC test.Conclusions SSD can cooperate with TMZ to inhibit the prolifera-tion of GBM cells and induce apoptosis and autophagy,and enhance the sensitivity of GBM cells to TMZ by ac-tivating endoplasmic reticulum stress pathway.

Immunotherapy is an important breakthrough in canc-er treatment.Unfortunately,low drug concentration in tumor sites almost ineffectively initiates immune responses and thereby severely limits immune therapy applications in clinics.Nanoma-terials are well-recognized drug delivery system in cancer thera-py.Nanographene oxide(NGO)have shown immense perti-nence for anti-cancer drug delivery owing to their ultra-high sur-face area,chemical stability,good biocompatibility and excel-lent photosensitivity.In addition,functionalized modifications on the surface of NGO increase tumor targeting and minimize cy-totoxicity.This study focuses on reviewing the literature and up-dates on NGO in drug delivery and discussing the possibilities and challenges of NGO in cancer synergetic therapy.

Aim To explore the effect of serum contai-ning Duhuo Jisheng Decoction on the proliferation,ap-optosis and inflammation of fibroblast-like synoviocytes(FLS)induced by tumor necrosis factor-α(TNF-α)in rheumatoid arthritis(RA)and to reveal the under-lying mechanism.Methods The MH7A cells were divided into five groups:normal group(10％blank se-rum),model group(10 μg·L-1 TNF-α+10％blank serum),Duhuo Jisheng Decoction low(10 μg·L-1 TNF-α+2.5％drug-containing serum+7.5％blank serum),medium(10 pg·L-1 TNF-α+5％drug-con-taining serum+5％blank serum),high(10 μg·L-1 TNF-α+10％drug-containing serum)dose group.The concentration of serum containing Duhuo Jisheng Decoction was screened by MTT method.Cell prolifer-ation was detected using EdU staining.The expression of proliferation marker Ki67 was detected using immu-nofluorescence staining.The apoptosis rate was meas-ured by flow cytometry.The contents of IFN-γ,IL-4,IL-6 and IL-10 in each group were detected by ELISA.The mRNA and protein expression of Bax,Bcl-2,caspase-3 and TLR4/MyD88/NF-κB signaling pathway were detected by RT-qPCR and Western blot.Results Compared with the normal group,the cell prolifera-tion activity of the model group significantly increased,and the level of apoptosis decreased.The content of IFN-γ and IL-6 increased,and the content of IL-4 and IL-10 decreased.The TLR4/MyD88/NF-κB signaling pathway was activated.After the intervention of low,medium and high dose groups of serum containing Du-huo Jisheng Decoction,it could effectively improve the abnormal proliferation of cells and enhance apoptosis.At the same time,it inhibited the inflammatory re-sponse and the activation of TLR4/MyD88/NF-κB sig-naling pathway.Conclusions The serum containing Duhuo Jisheng Decoction can inhibit the abnormal pro-liferation of RA-FLS induced by TNF-α and the secre-tion of pro-inflammatory factors,and enhance apoptosis and anti-inflammatory levels.The mechanism may be related to the regulation of TLR4/MyD88/NF-κB sig-naling pathway.

Aim To study the effect of salidroside combined with rosavin on ischemic stroke in rats.Methods The model of MCAO was established by u-sing thread-embolic method.The rats were divided into the sham group,MCAO group,salidroside combined with rosavin group,positive control group,and the drug was given continuously for seven days.The infarct volume was measured by MRI and neurological deficit score was evaluated by Zea-Longa.The levels of Ne-uN,BDNF,TGF-β1,p-Smad were observed by West-ern blot and immunofluorescence staining.The expres-sions of IL-1β,TNF-α and IL-6 were performed by RT-qPCR/ELISA.The primary cortical neurons were isolated,OGD/R inducted,divided into the normal group,OGD/R group,salidroside combined with rosa-vin group,and TGF-β1 inhibitor+salidroside com-bined with rosavin group,the drug was given for 24 hours,and the expressions of NeuN,BDNF,IL-1β,TNF-α and IL-6 were measured.Results Salidroside combined with rosavin could decrease the infarct vol-ume,improve the neurological function,promote the levels of Neun,BDNF,TGF-β1,p-Smad,and inhibit the expressions of IL-1β,TNF-α and IL-6.Salidroside combined with rosavin could promote NeuN,BDNF,inhibit IL-1β,TNF-α,IL-6 in primary nerve cells in-duced by OGD/R,and these effects were blocked by TGF-β1 inhibitor.Conclusions Salidroside combined with rosavin has neuroprotective effects on MCAO rats,and primary neurons are induced by OGD/R,and these effects are closely related to the TGF-β pathway.

Objective To compare the cost and utility of aflibercept and conbercept for the treatment of wet age-related macular degeneration(wetAMD),in order to provide a reference for the selection of treatment regimens from the perspective of pharmacoeconomics.Methods The incremental cost-utility ratio(ICUR)was obtained by using Markov model to simulate the survival of the two treatment regimens over the 5-year period,calculating costs and health outputs separately.Univariate sensitivity analysis was used to determine the impact of the parameter on ICUR,and probability sensitivity analysis was used to determine the influence of the uncertainty of each model parameter on the research results.One times the 2022 gross domestic product(GDP)per capita of China was used as the willingness-to-pay threshold(WTP)to judge its economy.Results Over the simulation period,the compazine regimen was significantly economical against the aflibercept regimen,with an ICUR of-1 528 840 per quality-adjusted life year(QALY),which was lower than the WTP.Univariate sensitivity analysis showed that the transition probability between mild and moderate visual status between the two regimens and the number of aflibercept injections per year were significant influencing factors of ICUR.Probabilistic sensitivity analysis pointed to a significant cost-utility advantage for conbercept at a WTP of one times GDP(probability of 65.9％),which was a more robust result.Conclusion For the treatment of wetAMD,conbercept has a cost-utility advantage compared with aflibercept.

Objective To compare the efficacy of single-port laparoscopic unilateral salpingectomy and adnexectomy(ovary+fallopian tube)via different approaches,and to explore the safety and feasibility of single-port single-handed laparoscopy without access platform.Methods A total of 94 patients who underwent unilateral salpingectomy and 78 patients who underwent unilateral adnexectomy in our hospital from August 2021 to November 2022 were selected and randomly divided into the single-port single-handed without access platform group,the single-port double-handed without access platform group,and the single-port with access platform group,respectively.The operation time,intraoperative blood loss,postoperative morbidity,postoperative anal exhaust time,postoperative hospitalization time,total hospitalization cost,incision length,postoperative pain visual analogue scale(VAS)score,surgical complications,postoperative body image scale(BIS),and cosmetic score(CS)of patients in the three groups were observed and compared.Results There was no statistically significant difference in operative time,intraoperative blood loss,postoperative morbidity,postoperative anal exhaust time,postoperative hospitalization time,and surgical complications among the three groups(P>0.05).There were statistically significant differences in total hospitalization cost,incision length,postoperative VAS score,postoperative BIS score,and postoperative CS among the three groups(P<0.05).Conclusion Single-port single-handed laparoscopy salpingectomy and adnexectomy without access platform is a safe and feasible surgical method,which has the advantages of smaller incision,less hospitalization cost,and higher incision satisfaction than single-port surgery with access platform and single-port double-handed laparoscopic surgery without access platform.

Objective To explore the expression of long non-coding ribonucleic acid(lncRNA)metastasis-associated lung adenocarcinoma transcript 1(MALAT1)and nuclear paraspeckle assembly transcript 1(NEAT1)in the hippocampus and HT22 cells of hypoxia pre-acclimated(HPC)mice and their relationship with neuroprotection.Methods(1)Thirty-six male Institute of Cancer Research(ICR)mice were randomly divided into three groups according to the random number table method of complete randomization:the control group,the hypoxia group and the hypoxia preconditioning group,with 12mice in each group.Mice in the control group were not exposed to hypoxia,mice in the hypoxia group were exposed to hypoxia once,and mice in the hypoxia preconditioning group were exposed to hypoxia four times.Immediately after the end of hypoxia treatment,all mice were decapitated and killed and hippocampal tissues were isolated and preserved in groups.(2)HT22 cells were cultured in medium containing 10％foetal bovine serum and 100 U/ml penicillin-streptomycin.When cell confluence was greater than 90％,they were transferred to 24-well plates for culture and then processed in 2 batches.6 pmol disordered small interfering RNA(siRNA),MALAT1 siRNA(siMALAT1),NEAT1 siRNA(siNEAT1),siMALAT1+siNEAT1 were transfected into the negative control group,siMALAT1 group,siNEAT1 group,and siMALAT1+siNEAT1 group of the first batch of HT22 cells one by one by transfection reagent,and the blank group did not have any treatment;then they were cultured under normal conditions(5％CO2 and 95％air)for 48 h.In the second batch of HT22 cells,6 pmol of disordered siRNA,disordered siRNA,siMALAT1,siMALAT1,siNEAT1 and siNEAT1 were transfected one by one correspondingly to the negative control group and the negative control+oxygen-glucose deprived/reoxygen(OGD/R)group,siMALAT1 group,siMALAT1+OGD/R,siNEAT1 group,siNEAT1+OGD/R group.48 h after transfection,HT22 cells of negative control group,siMALAT1 group and siNEAT1 group were cultured under normal conditions(5％CO2 and 95％air),and the cells of negative control+OGD/R group,siMALAT1+OGD/R group and siNEAT1+OGD/R group were treated with OGD/R.That is,under low oxygen conditions(1％O2+5％CO2+94％N2)exposure for 8 h,and then culture under normal conditions for 16 h.(3)The real-time fluorescence quantitative polymerase chain reaction(PCR)and Western blot was used to determine the expression of MALAT1,NEAT1,N-methyl-D-aspartate receptor subunit 2B(NR2B)messenger RNA(mRNA)and NR2B protein in the hippocampus of mice,the relative expression levels of NR2B mRNA and NR2B protein after transfection of HT22 cells in each group,and the relative expression levels of haemoglobin breakdown products and activated cysteine protease protein 3 after transfection and OGD/R of HT22 cells in each group.The survival rate of HT22 cells in each group was calculated.Results(1)The differences in relative expression of MALAT1(F=43.92),NEAT1(F=506.4),NR2B mRNA(F=50.64)and NR2B protein(F=41.24)in the hippocampus of mice in the three groups were statistically significant(all P＜0.05).The relative expression of MALAT1([1.68±0.06]vs.[1.00±0.08]),NR2B mRNA([1.26±0.06]vs.[1.00±0.01]),and NR2B protein([1.47±0.05]vs.[1.00±0.01])was increased in the hypoxia group as compared to the control group(all P＜0.05),whereas the relative expression of NEAT1([1.02±0.10]vs.[1.00±0.03])were not statistically significant(P＞0.05),and the relative expression of MALAT1([1.12±0.13]vs.[1.00±0.08])and NEAT1([2.88±0.10]vs.[1.00±0.03])were increased in hypoxic preconditioned group.Compared with hypoxia group,the relative expression of NR2B mRNA([0.54±0.07]vs.[1.26±0.06])and NR2B protein([1.17±0.07]vs.[1.47±0.05])were decreased(both P＜0.05).(2)The differences in the relative expression of NR2B mRNA(F=36.92)and NR2B protein(F=56.98)after transfection of HT22 cells in the five groups were statistically significant(both P＜0.05).Compared with the negative control group,siMALAT1 group(NR2B mRNA:[2.04±0.08]vs.[0.94±0.04],NR2B protein:[1.72±0.13]vs.[0.93±0.02]),siNEAT1 group(NR2B mRNA:[2.15±0.13]vs.[0.94±0.04],NR2B protein:[1.87±0.46]vs.[0.93±0.02]),siMALAT1+siNEAT1 group(NR2BmRNA:[2.09±0.16]vs.[0.94±0.04],NR2B protein:[2.07±0.30]vs.[0.93±0.02])showed the relative NR2B mRNA and NR2B protein expression were increased(all P＜0.05).(3)Differences in relative expression of haematopoietin breakdown product(145/150 kDa)protein(F=12.43),haematopoietin breakdown product(120 kDa)protein(F=7.15),and activated cysteamine protease protein 3 protein(F=6.61)were statistically significant in the 6 groups of HT22 cells transfected and treated with OGD/R(all P＜0.05).Compared with the siMALAT1 group,the siMALAT1+OGD/R group had 145/150kDa([1.42±0.48]vs.[0.85±0.34]),120 kDa([1.33±0.37]vs.[0.52±0.19])haematopoietin catabolism products and activated cysteamine protease protein 3([2.43±0.35]vs.[1.15±0.24])relative expression increased(all P＜0.05);compared with the negative control+OGD/R group,the siMALAT1+OGD/R group showed an increase in 145/150kDa([1.42±0.48]vs.[1.23±0.17]),120 kDa([1.33±0.37]vs.[0.80±0.21])relative expression of haematopoietin breakdown products and activated cysteamine protease protein 3([2.43±0.35]vs.[1.46±0.39])increased(all P＜0.05);compared with the siNEAT1 group,the siNEAT1+OGD/R group had a higher expression of 145/150 kDa([1.28±0.44]vs.[0.87±0.32]),120 kDa([0.81±0.36]vs.[0.63±0.16])relative expression of haematopoietic proteolytic products and activated cysteamine protease protein 3([1.51±0.45]vs.[1.01±0.27])increased(all P＜0.05).(4)The difference in HT22 cell survival rate among the 6 groups was statistically significant(F=5.54,P＜0.05).Compared with the negative control group,HT22 cell survival was decreased in the siMALAT1,siNEAT1,siMALAT1+OGD/R and siNEAT1+OGD/R groups([0.65±0.40],[0.76±0.35],[0.24±0.17],[0.23±0.16]vs.[0.84±0.04],all P＜0.05);cell viability was reduced in the siMALAT1+OGD/R group compared with the siMALAT1 group([0.24±0.17]vs.[0.65±0.40],P＜0.05);and cell viability was reduced in the siNEAT1+OGD/R group compared with the siNEAT1 group([0.23±0.16]vs.[0.76±0.35],P＜0.05).Conclusion HPC increased the expression of MALAT1 and NEAT1 in the hippocampus of mice,and MALAT1 and NEAT1 may participate in the neuroprotective effect of mice after ischemia and hypoxia by affecting the expression of NR2B.

Objective:To better evaluate the prevalence and epidemiological characteristics of metabolic-associated fatty liver disease (MAFLD) based on liver transient elastography.Methods:It was a cross-sectional study. A total of 6 961 patients without hepatitis, who underwent liver transient elastography examination at the Department of Hepatology, Zhongnan Hospital of Wuhan University from November, 2021 to April, 2022 were included. The patients were categorized into normal and mild, moderate, severe MAFLD groups according to FibroTouch controlled attenuation parameters (CAP). The CAP values among different body mass index (BMI) groups were compared using analysis of variance. The distribution characteristics and the incidence of MAFLD in different age, gender, body mass index(BMI), blood glucose and lipids groups using the chi-square test.Results:The total detection rate of MAFLD and severe MAFLD in the population with a BMI≥28.0 kg/m 2 was 99.6% and 71.8%, respectively. The detection rate of MAFLD in people with normal BMI was 28.4%. The detection rate of MAFLD in women of childbearing age or in the perimenopausal period were both significantly lower than that in men of the same age (40.3% vs 54.9%, χ 2=20.78, P<0.001; 43.1% vs 58.4%, χ 2=27.43, P<0.001), but there was no statistically significant difference in MAFLD detection rate between postmenopausal women and men of the same age. The detection rates of MAFLD in the group with abnormal blood glucose and lipids were both significantly higher than those in the group with normal blood glucose and lipids [69.7%(196/281) vs 35.2%(2 354/6 680), χ 2=138.36, P<0.001; 54.3%(1 696/3 124) vs 37.1%(1 420/3 837), χ 2=207.99, P<0.001]. Conclusion:Non-hepatitis patients had a higher prevalence of MAFLD. The BMI, gender, age, blood glucose, and lipids levels are all strongly associated with MAFLD.

Objective:To compare the recanalization rate of 2-suture longitudinal intussusception vasoepididymostomy(LIVE)with that of4-suture end-to-side vasoepididymostomy(ESVE).Methods:This retrospective case-control study included 127 cases of obstructive azoospermia(OA)treated by LIVE(n=87)or ESVE(n=40)in our Center of Reproductive Medicine from October 2013 to July 2024.We analyzed the clinical data and compared the age,testis volume,level of serum follicle-stimulating hormone(FSH),operation time and postoperative recanalization rate between the two groups.Results:Spermatozoa were observed in 90(70.9％)of the 127 cases after surgery.There were no statistical differences in age,testis volume and FSH between the two groups of patients(all P＞0.05),and nor were there any serious surgical complications.The operation time was significantly longer in the ESVE than in the LIVE group(22.0[20.0-25.8]vs 17.0[15.0-23.0]min,P＜0.05),while the postoperative recanalization rate remarkably higher in the former than in the latter group(85.0％vs 64.4％,P＜0.05).Vasoepididymostomy was performed at the caput epididymis in 11 cases,with a higher recanalization rate in the ESVE than in the LIVE group(50.0％[1/2]vs 33.3％[3/9]).Conclusion:ESVE achieved a higher postoperative recanalization rate than LIVE in the treatment of OA,but its advanta-ges need further investigation.

AIM To investigate the consistency of chemical constituents between formula granules and standard decoction of Coptidis Rhizoma.METHODS Eighteen batches of standard decoctions were prepared,after which the extraction rate and contents,transfer rates of magnolflorine,jatrorrhizine,columbamine,epiberberine,coptisine,palmatine,berberin were determined,HPLC characteristic chromatograms were established.RESULTS There were 11 common peaks in the characteristic chromatograms of 18 batches of standard decoctions and 24 batches of formula granules with the similarities of 0.861-1.000,which were clusterd into two categories.The formula granules and standard decoction demonstrated approximated extraction rate and contents,transfer rates of index constituents.CONCLUSION The chemical constituents between formula granules and standard decoction of Coptidis Rhizoma display good consistency,which can provide references for the quality control,process research and clinical application of the former.