Objective : To explore the expression profile of core 1 β1,3-galactosyltransferase 1(C1GALT1) in glioblastoma(GBM) and to elucidate its impact on the initiation and progression of GBM. Methods : The expression levels and prognostic significance of C1GALT1 in GBM were analyzed using the GEPIA and CGGA databases. Two representative glioblastoma cells(U251 and LN18) were selected to construct C1GALT1-knockdown cell lines and performed in vitro experiments. The Cell Counting Kit-8(CCK-8) and Transwell assays were employed to evaluate the impact of C1GALT1 on proliferation, migration and invasion of GBM cells. Transcriptome data were analyzed to identify potential signaling pathways. Senescence β-Galactosidase Staining Kit was used to detect β-galactosidase activity. Results : nalysis of GEPIA and CGGA databases revealed that C1GALT1 was significantly upregulated in GBM tissues compared to adjacent non-cancerous tissues (P < 0. 05) , and its high expression was associated with poor prognosis of patients (P < 0. 000 1) . The CCK-8 experiment demonstrated a significant reduction in prolifera- tion rate following C1GALT1 knockdown (P < 0. 05) . Transwell assay showed that cell migration and invasion de- creased after C1GALT1 was knocked down ( P < 0. 001) . Transcriptome sequencing and senescence β-galactosi- dase staining showed that C1GALT1 was involved in the cellular senescence signaling pathway , and the activity of β-galactosidase associated with cellular senescence significantly increased after C1GALT1 was knocked down(P < 0. 05) . Conclusion C1GALT1 is overexpressed in GBM tissues and may promote the proliferation , migration and invasion of GBM cells by inhibiting cellular senescence .

Objective:To explore the impact of Epstein-Barr Virus(EBV) infection on treatment response and survival in newly diagnosed multiple myeloma(MM).Methods:The clinical data of 196 patients with newly diagnosed MM admitted to Zhongshan Hospital of Fudan University from June 1st, 2019 to February 25th,2021 were analyzed retrospectively and divided into EBV-positive group (106 cases) and negative group (90 cases) according to the primary EBV DNA results in peripheral blood mononuclear cells.To analyse the distribution of EBV positive rates in each type and in each stage of the Revised International Staging System (R-ISS), and to compare EBV DNA loads in EBV-positive patients among R-ISS stages.Rank sum test, 2×2 chi-square test and independent sample t-test were used to compare laboratory findings, such as liver and kidney function, immunohistochemistry and cytogenetics, treatment efficacy and survival prognosis between the two groups.The clinical prognosis of EBV-positive patients was summarized through survival analysis and Cox regression.Results:The EBV positive rate in patients with newly diagnosed MM was 54% (106/196), with the highest rate in patients with κ light chain type (9/12).Patients with R-ISS stage Ⅲ had a significantly higher positive rate than with stage Ⅰ ( χ2=4.68, P=0.031) and stage Ⅱ ( χ2=6.04, P=0.014), but there was no significant difference in EBV DNA loads between EBV-positive MM patients by stage ( Z=3.27, P=0.195).Serum creatinine (Scr) and β 2-microglobulin (β 2-MG) levels were higher in the EBV-positive group than in the EBV-negative group ( Z=1.98, P=0.048 and Z=2.08, P=0.038), and the occurrence of t(4;14) was also higher in the EBV-positive group ( χ2=3.93, P=0.047).The proportion of complete response (CR)/stringent complete response(sCR) and very good partial response(VGPR) after completion of the fourth chemotherapy were significantly lower in the EBV-positive group than in the EBV-negative group ( χ2=12.82, P=0.001 and χ2=8.30, P=0.004), and a higher rate of progressive disease (PD) occurred in the EBV-positive group ( χ2=4.48, P=0.046).The 2-year progression-free survival (PFS) of MM patients was shorter in the EBV-positive group compared to that in the EBV-negative group ( Z=-4.50, P0.01).Cox regression analysis showed that R-ISS stage Ⅲ ( HR=5.38, 95% CI 1.28-22.56, P=0.021), failure to achieve VGPR after the fourth chemotherapy ( HR=3.02, 95% CI 1.42-6.46, P=0.004), EBV-positive ( HR=1.98, 95% CI 1.02-3.87, P=0.045), with 1q21 amplification ( HR=2.35, 95% CI 1.16-4.75, P=0.017) and 13q14 deletion ( HR=1.93, 95% CI 1.01-3.67, P=0.046) were independent risk factors for PFS in newly diagnosed MM. Conclusions:EBV infection is an independent risk factor for poor prognosis, which has important clinical implications for the outcome and prognosis of patients with newly diagnosed MM, and may become a novel clinical assessment indicator.

Objective:To explore the impact of Epstein-Barr Virus(EBV) infection on treatment response and survival in newly diagnosed multiple myeloma(MM).Methods:The clinical data of 196 patients with newly diagnosed MM admitted to Zhongshan Hospital of Fudan University from June 1st, 2019 to February 25th,2021 were analyzed retrospectively and divided into EBV-positive group (106 cases) and negative group (90 cases) according to the primary EBV DNA results in peripheral blood mononuclear cells.To analyse the distribution of EBV positive rates in each type and in each stage of the Revised International Staging System (R-ISS), and to compare EBV DNA loads in EBV-positive patients among R-ISS stages.Rank sum test, 2×2 chi-square test and independent sample t-test were used to compare laboratory findings, such as liver and kidney function, immunohistochemistry and cytogenetics, treatment efficacy and survival prognosis between the two groups.The clinical prognosis of EBV-positive patients was summarized through survival analysis and Cox regression.Results:The EBV positive rate in patients with newly diagnosed MM was 54% (106/196), with the highest rate in patients with κ light chain type (9/12).Patients with R-ISS stage Ⅲ had a significantly higher positive rate than with stage Ⅰ ( χ2=4.68, P=0.031) and stage Ⅱ ( χ2=6.04, P=0.014), but there was no significant difference in EBV DNA loads between EBV-positive MM patients by stage ( Z=3.27, P=0.195).Serum creatinine (Scr) and β 2-microglobulin (β 2-MG) levels were higher in the EBV-positive group than in the EBV-negative group ( Z=1.98, P=0.048 and Z=2.08, P=0.038), and the occurrence of t(4;14) was also higher in the EBV-positive group ( χ2=3.93, P=0.047).The proportion of complete response (CR)/stringent complete response(sCR) and very good partial response(VGPR) after completion of the fourth chemotherapy were significantly lower in the EBV-positive group than in the EBV-negative group ( χ2=12.82, P=0.001 and χ2=8.30, P=0.004), and a higher rate of progressive disease (PD) occurred in the EBV-positive group ( χ2=4.48, P=0.046).The 2-year progression-free survival (PFS) of MM patients was shorter in the EBV-positive group compared to that in the EBV-negative group ( Z=-4.50, P0.01).Cox regression analysis showed that R-ISS stage Ⅲ ( HR=5.38, 95% CI 1.28-22.56, P=0.021), failure to achieve VGPR after the fourth chemotherapy ( HR=3.02, 95% CI 1.42-6.46, P=0.004), EBV-positive ( HR=1.98, 95% CI 1.02-3.87, P=0.045), with 1q21 amplification ( HR=2.35, 95% CI 1.16-4.75, P=0.017) and 13q14 deletion ( HR=1.93, 95% CI 1.01-3.67, P=0.046) were independent risk factors for PFS in newly diagnosed MM. Conclusions:EBV infection is an independent risk factor for poor prognosis, which has important clinical implications for the outcome and prognosis of patients with newly diagnosed MM, and may become a novel clinical assessment indicator.

Objective To investigate the expression of miR-27a in colorectal cancer cell,and to analyze the effect of its targeted regulation of(Secreted Frizzled-Related Protein,SFRP1)on the biological behavior of colorectal cancer cells.Methods Real-time fluorescent quantitative PCR(qRT-PCR)was employed to examine the expres-sion of miR-27a and SFRP1 mRNA in colorectal cancer tissues and adjacent normal tissues.Western blot was used to detect the expression of SFRP1 protein in colorectal cancer tissues and adjacent normal tissues.TargetScan soft-ware and dual luciferase reporter gene test were used to detect the targeted regulation of miR-27a on SFRP1.HCT116 cells were transfected with miR-27a mimic,miR-27a inhibitor and negtive control(NC).The expression of miR-27a and SFRP1 mRNA in each group was determined by qRT-PCR.MTT colorimetry was performed to eval-uate the proliferation of each group cells.Transwell assay was used to evaluate the cell invasion and migration abili-ty.Meanwhile,the protein expression levels of SFRP1,key factors Wnt4 and β-catenin in the Wnt/β-catenin sig-naling pathway were determined by Western blot.Results Compared with adjacent normal tissues,miR-27a was highly expressed in colorectal cancer tissues,while SFRP1 was low expressed in colorectal cancer tissues(P<0.05).TargetScan software and dual luciferase reporter gene test showed that miR-27a targeted SFRP1.Compared with NC group,the expression of miR-27a of miR-27a mimic group increased,the proliferation,invasion and mi-gration ability enhanced,the expression of SFRP1 protein decreased,while Wnt4 and β-catenin protein expression increased(P<0.05).Compared with miR-27a mimic group,the expression of miR-27a of miR-27a inhibitor group decreased,the proliferation,invasion and migration ability reduced,the expression of SFRP1 protein in-creased,while Wnt4 and β-catenin protein expression decreased(P<0.05).Conclusion miR-27a can target SFRP1,inhibit the proliferation,invasion and migration of colorectal cancer cells,mainly by up-regulating SFRP1 and blocking the downstream Wnt/β-catenin signaling pathway,which provides a new direction for clinical treat-ment.

Objective To investigate the effect of aspirin-induced Gasdermin E(GSDME)dependent pyroptosis and the related mechanism of colorectal cancer cell proliferation.Methods The colorectal cancer cells Caco2 were cultured in vitro.MTT assay was used to detect the effects of aspirin intervention with different concentrations(1.0,2.5,5.0,10.0,15.0,20.0 mmol/L)on the proliferation activity of colorectal cancer cells.The effect of aspirin intervention on the morphology of colorectal cancer cells was observed under the microscope.Lactate dehy-drogenase(LDH)release assay was used to investigate the effect of aspirin intervention on cell membrane integrity.The protein expression levels of NOD-like receptor protein(NLRP3),cysteinyl aspartate and specific proteinase 1(Caspase-1),Gasdermin E-N(GSDME-N)in colorectal cancer cells were detected by Western blot.The contents of interleukin-1 β(IL-1 β)and interleukin-18(IL-18)in cell supernatant after GSDME silencing were detected by ELISA.After silencing GSDME,cell morphological changes were observed under a microscope,and cell membrane integrity was observed by LDH release assay;The contents of IL-1 β and IL-18 in cell supernatant were determined by ELISA after GSDME silencing.Results MTT results showed that aspirin could decrease the proliferation activi-ty of Caco-2 in a concentration-dependent manner(P＜0.01).Morphological observation and LDH experiment showed that aspirin could promote the occurrence of pyroptosis(P＜0.05).Western blot showed that aspirin could increase the expression levels of NLRP3,Caspase-1,GSDME-N of pyroptosis-related pathway genes(P＜0.05).ELISA showed that aspirin could significantly increase the concentration of IL-1 β and IL-18 in Caco-2 cells(P＜0.01).After GSDME silencing,the pyroptosis was significantly inhibited(P＜0.05)and the expression of IL-1 βand IL-18 decreased significantly(P＜0.01).Conclusion Aspirin can inhibit the proliferation of Caco-2 by indu-cing the pyroptosis of GSDME-dependent cells,thus inhibiting colon cancer.

OBJECTIVE To establish a method for simultaneous determination of zeaxanthin ，β-carotene，β-cryptoxanthin palmitate and zeaxanthin dipalmitate in Lycium barbarum . METHODS L. barbarum was extracted with n-hexane-anhydrous ethanol-acetone-toluene（10∶6∶7∶7，V/V/V/V）by ultrasonic method. High performance liquid chromatography （HPLC）method was adopted. The determination was performed on YMC C 30 column with mobile phase A consisted of methanol-acetonitrile-water （81∶ 14 ∶ 5，V/V/V）and mobile phase B consisted of dichloromethane （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 20 ℃. The detection wavelength was set at 450 nm，and sample size was 20 μL. Using zeaxanthin as control，the relative correction factors （RCFs）of β-carotene，β-cryptoxanthin palmitate and zeaxanthin dipalmitate were calculated ， and then the content of each component was calculated according to RCFs and compared with the results of external standard method（ESM）. RESULTS The linear range of zeaxanthin ，β-carotene，β-cryptoxanthin palmitate and zeaxanthin dipalmitate were 0.119 4-2.983 8，0.121 7-1.521 6，0.285 9-5.718 8，8.460 5-211.513 3 μg/mL（all R2＞0.999）. RSDs of precision ，repeatability and stability（16 h）tests were all less than 4％. The average recoveries were 103.34％，107.37％，105.64％，96.16％（RSD＜5％，n＝ 9）. The average RCFs of β-carotene，β-cryptoxanthin palmitate and zeaxanthin dipalmitate were 1.109，1.390，1.158. The relative errors of the content determination results by quantitative analysis of multi-components by singer marker （QAMS）and ESM were within ±1％. CONCLUSIONS The established HPLC-QAMS method is accurate and stable ，which can be used for the content determination and quality control of 4 carotenoids in L. barbarum .

Objective To analezk thk charactkristics of drug-induckd livkr injure( DIFI)in childrkn with acutk lemphoblastic lkuckmia(LFF),so as to improvk thk phesician＇s undkrstanding of chkmothkrape DIFI,and to guidk clinical rational drug usk. Methods Onk hundrkd and forte-thrkk casks with LFF diagnoskd in thk Dkpartmknt of Hk-matologe and Oncologe in thk Pirst Lffiliatkd Hospital of Yhkngzhou Rnivkrsite from Januare 2012 to Dkckmbkr 2016 wkrk analezkd rktrospkctivkle. Baskd on DIFI diagnostic critkria and thk ARCLM scalk,thk casks with a scork of ≥3 points wkrk considkrkd to havk chkmothkrape DIFI. Groupkd be gkndkr,agk,immunoteping,risc and stagk of chkmo-thkrape,thk incidknck of DIFI was comparkd. Thk situation aftkr DIFI prkvkntion was comparkd bktwkkn two groups which was groupkd according to whkthkr thk application of hkpatoprotkctivk drugs. ResuIts Onk hundrkd and kight ca-sks(75. 52﹪)had DIFI,66 casks(61. 11﹪)showkd clinical manifkstations of livkr injure,and 42 casks(38. 89﹪) had no clinical semptoms. Lmong all thk casks 57. 41﹪(62 casks)wkrk mild livkr damagk,25﹪(27 casks)wkrk modkratk livkr injure and 17. 59﹪(19 casks)wkrk skvkrk livkr damagk. Thk clinical tepks which wkrk hkpatockllular accounting for 79. 63﹪(86 casks),cholkstatic 7. 41﹪(8 casks)and mixkd 12. 96﹪(14 casks). Malk wkrk 80 casks (79. 21﹪)and fkmalk 28 casks(66. 67﹪),but thk incidknck of DIFI bktwkkn diffkrknt gkndkr group had no statistical diffkrknck(χ2 ﹦2. 524,P﹦0. 112). Skvknte-fivk casks(77. 32﹪)wkrk ＜7 ekars agk and 33 casks(71. 74﹪)≥7 ekars agk,and thk incidknck of DIFI bktwkkn 2 groups was not statisticalle diffkrknt(χ2 ﹦0. 526,P﹦0. 468). Thkrk was no significant diffkrknck in T-LFF(8 casks,61. 54﹪)and B-LFF(100 casks,76. 92﹪)( χ2 ﹦0. 795,P﹦0. 372). Thk incidknck had significant diffkrknck in diffkrknt risc(P﹦0. 002). Thk incidknck of DIFI bktwkkn thk middlk risc group(60 casks,88. 24﹪)and standard risc(21 casks,58. 33﹪)had statistical diffkrknck( P ＜0. 05 ). Thk incidknck of DIFI bktwkkn thk middlk risc group and skvkrk risc(27 casks,69. 23﹪)had statistical diffkrknck( P﹦0. 015). Thk incidknck was diffkrknt in diffkrknt stagks of chkmothkrape(P＜0. 05). Thk incidknck of DIFI in induckd stagk was diffkrknt comparkd to othkr stagks(P＜0. 05). ARCLM scork ＞8 points accountkd for 21 casks(19. 45﹪), 6-8 points accountkd for 59 casks(54. 63﹪)and 3 -5 points accountkd for 28 casks(25. 92﹪). Eighte -nink patiknts(92. 71﹪)wkrk kffkctivk in thk hkpatoprotkctivk group and 8 patiknts(66. 67﹪)in thk no hkpatoprotkctivk thkrape group. Thk diffkrknck bktwkkn thk 2 groups was statisticalle significant(χ2 ﹦5. 317,P﹦0. 021). ConcIusions Thk clinical semptoms of drug-induckd livkr injure in childrkn with LFF chkmothkrape ark lacc of spkcificite. Thke ark mainle charactkrizkd be mild livkr injure. Thk clinical tepk of hkpatic injure is common in hkpatockllular. Thk ARCLM scork was mostle 6 to 8. Thkrk is no rklationship bktwkkn thk incidknck in LFF and gkndkr,agk,tepk of lkuck-mia. Thk incidknck with modkratk risc tepk is highkr than that of thk standard and high-risc tepk. Thk incidknck in induction rkmission stagk is highkst. Lpplication of hkpatoprotkctivk drugs is bknkficial to DIFI prognosis.

Deep brain disease stimulation (DBS) is commonly used to treat movement disorders such as Parkinson disease,and current evidence suggests that DBS may also be useful for refractory epilepsy and is affected by a variety of factors.Studies show that stimulation of the anterior nucleus of the thalamus and hippocampus may decrease the frequency of refractory seizures.The efficacy of stimulating other targets remains inconclusive.An absence of structural abnormality on imaging and electrode position are associated with stimulation efficacy.Certain seizure types may respond more favorably to specific targets.There are several factors that potentially predict seizure outcome following DBS,but more large-scale clinical trials are needed.

Objective To evaluate the effect of atorvastatin preconditioning on cognitive function in isoflurane-anaesthetized mice.Methods Forty-eight healthy male C57BL/6 mice,aged 3 months,weighing 27-41 g,were divided into 3 groups (n =16 each) using a random number table:control group (group C),isoflurane anesthesia group (group Ⅰ) and atorvastatin preconditioning plus isoflurane anesthesia group (group AI).Atorvastatin 10 mg/kg was given through a gastric tube into the stomach at the same time every day for 7 consecutive days in group AI.In Ⅰ and AI groups,1.5％ isoflurane was inhaled for 6 h with fresh gas flow of 2 L/min at 1 day after the end of administration.Open field test and Morris water maze test were performed at 1 day after the end of anesthesia.The mice were sacrificed at 1 day after the end of Morris water maze test,and hippocampi were isolated for determination of caspase-3,Bax and Bcl-2 expression (by Western blot) and contents of interleukin-1beta (IL-1β),tumor necrosis factor-alpha (TNF-α) and soluble Aβ1-42 in hippocampal tissues (by enzyme-linked immunosorbent assay).Results There was no significant difference in the parameters of open field test among the three groups (P＞0.05).Compared with group C,the escape latency was significantly prolonged at each time point,the time of staying at the original platform quadrant was shortened,the frequency of crossing the original platform was decreased,the contents of IL-1β,TNF-α and soluble Aβ1-42 were increased,the expression of caspase-3 and Bax was up-regulated,and Bcl-2 expression was down-regulated in Ⅰ and AI groups (P＜0.05).Compared with group Ⅰ,the escape latency was significantly shortened at each time point,the time of staying at the original platform quadrant was prolonged,the frequency of crossing the original platform was increased,the contents of IL-1β,TNF-α and soluble Aβ1-42 were decreased,the expression of caspase-3 and Bax was downregulated,and Bcl-2 expression was up-regulated in group AI (P＜0.05).Conclusion Atorvastatin preconditioning can improve cognitive function in isoflurane-anaesthetized mice,and the mechanism may be association with attenuating hippocampal inflammatory responses,inhibiting over-expression of Aβ1-42 and inhibiting neuronal apoptosis.

Objective To investigate the effects of Trim34α on the activation of luciferase reporter gene containing NF-κB promoter induced by adaptor proteins TAB2. Methods The total RNA was isolated from HeLa cells. After amplification with RT-PCR, the target sequences were cloned into 5'-Flag-pcDNA3.1 (+) vector. The recombinant vector was confirmed by restriction enzyme digestion, colony PCR and sequencing. It was transfected into HEK293T cells to detected Trim34α expression by Western blot. Simultaneously, the effects of Trim34α on the NF-κB activation induced by TAB2 were determined by dual-luciferase reporter assay. Results Restriction enzyme digestion, colony PCR and sequencing confirmed the vector was constructed successfully, furthermore it expressed Trim34α protein in HEK293T cells. Moreover, trim34α could form high-molecular-weight oligomeric protein, and here we called it trimsome. Interestingly, dual-luciferase assay showed that Trim34α could effectively block TAB2-induced NF-κB activation. Conclusion Trim34α was involved in negative regulation of TAB2-induced NF-κB activation and could form high-molecular-weight oligomer.