Objective To study the radiosensitivity enhancement effects of recombinant adenovirus mediated retinoblastoma 94 gene on the growth of esophageal carcinoma cells EC109.Methods EC109cells was transfected with recombinant Rb94 gene adenovirus and irradiated by 137Cs γ-rays.The cohorts were divided into groups as blank control,Ad-LacZ,Ad-Rb94,radiation and Ad-Rb94 combined with radiation.Cell inhibition ratio,cell cycle and expression of retinoblastoma protein of EC109 cells were analyzed.Results The growth of EC109 cells transfected with Ad-Rb94,radiation and Ad-Rb94 combined with radiation group was all inhibited.The group of Ad-Rb94 combined with radiation resulted in greater inhibition of cells growth compared with Ad-Rb94 group and radiation group ( F =23.31,P ＜0.05 ).Cells of G2 phases of EC109 cancinoma cells for Ad-Rb94 combined with radiation group were the highest,which was 50％.The combination of Ad-Rb94 and radiation group resulted in the greatest expression of retinoblastoma protein,which reached 71％,significantly higher than Ad-Rb94 infection and radiation groups ( x2 =8.31,6.73,P ＜ 0.05 ).Conclusions Retinoblastoma 94 gene combined with ionizing radiation can enhance the radiation sensitivity of EC109 cells.

Objective To prepare a human meningococcal reference serum and standardize IgG concentrations to capsular polysaccharides and in vitro bactericidal activities of the reference serum against serogroup A, C, Y and W135 strains.Methods Twenty healthy adults were recruited and given one dose of immunization with tetravalent (serogroups A, C, Y and W135) meningococcal polysaccharide vaccine . Plasma samples were collected and gone through a series of process treatments including defibrination , filtra-tion, and lyophilization to prepare the meningococcal reference serum Men 10.The IgG concentrations of Men10 to capsular polysaccharides of serogroups A , C, Y and W135 were calibrated by using an internation-al reference serum CDC1992 as the standard in enzyme-linked immunosorbent assay (ELISA).Provisional IgG concentrations of Men10 were intensively validated by testing a panel of 12 calibration serum samples from Centers for Disease Control and Prevention , USA ( US CDC) and a panel of 56 serum samples immu-nized with A, C, Y and W135 meningococcal polysaccharide vaccine from Lanzhou Institute of Biological Products Co., Ltd.(LIBP) with the assays using Men10 and CDC1992 as the standard and/or test sam-ples, respectively.The bactericidal titers against serogroup A , C, Y and W135 strains were measured by se-rum bactericidal assay (SBA).Results Four thousand vials (0.5 ml/vial)of lyophilized human meningo-coccal reference serum Men10 were successfully prepared with 2.5%of residual moisture .Reference serum Men10 was sterile and free from contamination by hepatitis B virus , hepatitis C virus , human immunodefi-ciency virus and syphilis .Provisional IgG concentration of Men 10 to capsular polysaccharide of serogroups A, C, Y and W135 was calibrated by using CDC1992 as the standard.Furthermore, IgG concentrations of both panels of 12 CDC calibration serum samples and 56 LIBP serum samples calibrated by using Men 10 as the standard correlated well with those by using CDC1992 as the standard (r=0.99,P<0.05).The IgG concentrations of CDC1992 as calibrated by using Men10 as the standard showed significant correlation with its previously determined values with variation <10%.SBA titers for serotype A , C, Y and W135 strains were established as well .Conclusion A panel of new human meningococcal reference serum Men 10 with accurately calibrated IgG concentration against capsular polysaccharide of serogroups A , C, Y and W135 as well as SBA titers was successfully established .

ObjectiveTo investigate the effects of maltol aluminum exposure on miR-193a-3p, demethylase AlkB homolog 5 (ALKBH5), phosphatase and tensin homolog deleted on chromosome ten (PTEN) and protein kinase B (AKT), and whether miR-193a-3p is involved in aluminum-induced cognitive impairment by regulating ALKBH5/PTEN/AKT signaling pathway. Methods Specific pathogen-free male SD rats were randomly divided into control group and low-, medium- and high- dose groups according to their body weight, with eight rats in each group. Rats in the low-, medium-, and high- dose groups were intraperitoneally injected with maltol aluminum solution at concentrations of 10.00, 20.00, and 40.00 μmol/kg body weight, respectively, while the rats in control group were given an equal volume of 0.9% sodium chloride solution. Rats were injected for five days every week for three months. After injection, the novel object recognized test was used to assess the learning and memory ability of the rats. The relative expression of miR-193a-3p and B-cell lymphocytoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cysteine aspartate protease-3 (Caspase-3) mRNA in rat hippocampus was detected using the real-time quantitative polymerase chain reaction. The relative protein expression of ALKBH5, PTEN, and AKT2 in the rat hippocampus was detected using Western blot. Results The discrimination index and the preference index of the new object recognition test of the rats in high-dose group were lower than those in control group and low-dose group (all P<0.05). The relative expression of miR-193a-3p and Bcl-2 mRNA in the hippocampus of the rats in high-dose group was lower than those in control group and low-dose group (all P<0.05). The relative mRNA expression of Bax in the high-dose group was higher than those in the control group and low-dose group (both P<0.05). The relative mRNA expression of Caspase-3 of the rats in the high-dose group was higher than that in the other three groups (both P<0.05). The relative protein expression of ALKBH5 in the hippocampus of the rats in the high-dose group was lower than that in the control group (P<0.05). The relative expression of PTEN protein was higher than those in the control group and low-dose group (both P<0.05). The relative protein expression of AKT2 was lower than those in the control group and low-dose group (both P<0.05). Conclusion Sub-chronic aluminum exposure can inhibit the expression of miR-193a-3p in the hippocampus of rats, which may disrupt the ALKBH5/PTEN/AKT pathway and affect normal neuronal homeostasis and cellular function. This pathway may play an important role in aluminum-induced cognitive impairment.

The cartilage-protective effect of ginkgolide C(GC)on the two modeling modalities was investigated based on joint pain,degree of cartilage pathology,ECM degradation process,and level of inflammatory mediator production in rats.Twenty-five SD rats were selected and randomly di-vided into five groups:the control group(Control group),model 1 group(ACLT group),adminis-tration 1 group(ACLT+GC group),model 2 group(MIA group),and administration 2 group(MIA+GC group.)The rats were euthanized after 4 weeks of the test.Femur,tibia and blood samples were collected from the right hind limb of rats.The degree of pathology in the femur and tibia of rats was assessed by saffron O solid green staining and OARSI score.Immunohistochemis-try was used to detect the expression levels of collagen Ⅱ and MMP-13 in cartilage.ELISA was used to detect the changes in the levels of MMP-3,MMP-13,CTX-Ⅱ,COMP,COX-2,INOS,IL-1β,and TNF-α in the serum of rats.Cold sensitivity test and knee extension vocalization test were conducted to detect the degree of joint pain in rats.ACLT could cause more severe structural dam-age to articular cartilage compared with the MIA group.The OARSI scores and the expression of MMP-13 in femur and tibia,and the serum levels of MMP-13,MMP-3,CTX-Ⅱ,and COMP were higher in the ACLT group than those in the MIA group.However,the levels of inflammatory me-diators COX-2,IL-1β,and TNF-α were significantly lower in the ACLT group than in the MIA group(P＜0.0l).GC intervention reduced the OARSI score(P＜0.05 or P＜0.01)and pain scores,inhibited the ECM matrix degrading enzymes(MMP-13,MMP-3),cartilage metabolism markers(CTX-11,COMP),and inflammatory mediators(COX-2,INOS,IL-1β and TNF-α)ex-pression,and promoted collagen Ⅱ synthesis.Both modeling methods resulted in cartilage damage.In particular,the OA model constructed by ACLT+PMMx method in rats had obvious joint dam-age,which was favorable to investigate the degree of cartilage structural damage.GC attenuated cartilage pathological changes,pain severity and inflammatory response in the rat OA model in both groups,thus exerting a cartilage-protective effect.

AIM:To investigate the effects of subchronic aluminum exposure on the expression of silent infor-mation regulator(Sirt1),Kelch-like ECH-associated protein 1(Keap1),nuclear factor E2-related factor 2(Nrf2),and microRNA-128-3p(miR-128-3p)in the hippocampus of rats.Additionally,we aimed to explore the mechanism of miR-128-3p and the Sirt1-Keap1/Nrf2 signaling pathways in aluminum-induced cognitive impairment in rats.METHODS:Thirty-two healthy 6-week-old SPF male SD rats,weighing(190±20)g,were randomly divided into four groups based on body weight:control group,low-dose(10 μmol/kg)group,medium-dose(20 μmol/kg)group,and high-dose(40 μmol/kg)group,with 8 rats in each group.The rat exposure model was established by intraperitoneal injection of maltol alumi-num.The Morris water maze test was used to assess the learning and memory ability of the rats.Western blot analysis was performed to measure the protein expression of Sirt1,Keap1 and Nrf2 in the hippocampus,while RT-qPCR was used to measure the expression of miR-128-3p in the hippocampus.The level of reactive oxygen species(ROS)in the cerebral cor-tex was detected using fluorescence staining in frozen sections.RESULTS:(1)In the positioning cruise experiment,the escape latency of the aluminum exposure group was significantly higher than that of the control group on the 3rd,4th,and 5th days(P＜0.05).On day 6,the number of times the rats crossed the platform and the platform quadrant in the high-dose group was reduced compared to the control and low-dose groups(P＜0.01).(2)The expression levels of Sirt1 and Nrf2 in the hippocampal tissues of all groups decreased gradually with increasing maltol aluminum exposure dose.The ex-pression level of Keap1 increased gradually with increasing maltol aluminum exposure dose.The expression level of miR-128-3p in the high-dose group was significantly higher than that in the control group(P＜0.05).(3)The content of gluta-thione peroxidase in the hippocampus of rats decreased with increasing exposure dose,while ROS levels gradually in-creased.CONCLUSION:Subchronic aluminum exposure can increase the expression of miR-128-3p in the rat hippo-campus and suppress the Sirt1-Keap1/Nrf2 signaling pathway.This inhibition prevents the activation of the Sirt1-Keap1/Nrf2 signaling pathway,leading to a reduced antioxidant capacity.The imbalance in the antioxidant system in rats results in oxidative damage to nerve cells and a subsequent decline in cognitive function.

Purpose To explore the ability of proton density fat fraction(PDFF)and decay constant T2* values in MRI to reflect skeletal muscle aging.Materials and Methods 3T MRI data of skeletal muscle in the middle thigh of 211 healthy adults from the Third Affiliated Hospital of Southern Medical University from August to December 2023 were prospectively collected.Gender,age,height,weight and body mass index(BMI)were recorded.PDFF value and T2* value of thigh skeletal muscle were measured at post-processing workstation,and statistical differences among different age,gender and BMI groups were analyzed.The correlation between PDFF value and T2* value of thigh skeletal muscle and age and BMI was analyzed.Results There were statistically significant differences in PDFF values of thigh skeletal muscle among different age groups(H=18.476-85.619,all P<0.01).There were significantly differences in T2*values of the left and right quadriceps muscles,hamstrings and adductors among different age groups(H=13.342-47.566,all P<0.05).There were statistically significant differences in the PDFF values of right quadriceps,left and right hamstring,adductor and sartor muscles between male and female groups(Z=-4.929--1.626,all P<0.05),while there were statistically significant differences in T2* values of left sartor muscle(Z=-2.971,P=0.003).There was no statistical significance in PDFF value of skeletal muscle of thigh in different BMI groups(P>0.05),but there were statistically significant differences in T2* value of left and right quadriceps muscle,hamstring muscle and adductor muscle(H=9.542-24.495,all P<0.05).There was a moderate positive correlation between age and PDFF value of thigh skeletal muscle(r=0.635,P<0.01),but a slight negative correlation with T2* value of left and right quadriceps,hamstring and sarcoleus(r=-0.451--0.189,all P<0.01).There was a slight positive correlation between BMI and T2* values of thigh skeletal muscle(r=0.317,P<0.01).There was a moderate negative correlation between the PDFF value and T2* value of all thigh skeletal muscles(r=-0.749--0.624,P<0.01).The PDFF and T2* values of the front and back thigh muscles(quadriceps,hamstring)were most significantly correlated with age and BMI.Conclusion PDFF based on MRI can reflect the age-related changes in the microenvironment of thigh skeletal muscle,and is a potential imaging biological marker for accurate and non-invasive quantitative evaluation of thigh skeletal muscle aging.

Objective To investigate maternal zinc metabolism and the changes of zinc-related factors as metallothionein-1 (MT1) and zinc transporter-1 (ZnT1) in certain types of congenital heart diseases (CHD).Methods Fifteen infants with interventricular septal defect,12 infants with atrial septal defect and 7 infants with tetralogy of Fallot,together with their mothers were enrolled,and normal infants and their mothers were enrolled by a ratio of 1 ∶ 1 with the above three types of CHD diseases.General conditions of the mothers,along with their diets and zinc-containing drug supplementation during the pregnancy,were surveyed.Maternal blood zinc levels and serum alkaline phosphatase activities at gestation week 32 and delivery or induced abortion,and the protein and mRNA expressions of MT1 and ZnT1 in maternal serum and placental tissue at delivery or induced abortion were assayed.Results The general conditions were comparable between the CHD group and control group.The ratio of the mothers taking more zinc-rich food was significantly lower in the CHD group than in the control group.Circulating zinc levels in interventricular septal defect (73.55±5.79 μmol/L),atrial septal defect (72.66±5.82 μmol/L) and tetralogy of Fallot (68.72±6.72 μmol/L) groups were significantly lower than those in the control groups (82.77± 7.88,84.58 ± 7.55 and 85.66 ± 7.30 μmol/L) at delivery (P all ＜ 0.05).Similar change patterns were seen for serum alkaline phosphatase activities.The relative quantities of serum MT1 and ZnT1 proteins in interventricular septal defect (73.22±36.54 and 68.55± 27.82),atrial septal defect (64.29± 38.26 and 74.55 ± 29.67) and tetralogy of Fallot (67.88± 30.50 and 70.13±29.65) groups were significantly lower than those in their corresponding control groups (166.31±67.43and 97.67±30.22,182.56±71.40 and 111.65±32.70,and 173.81±62.36 and 108.27±28.52,P＜0.01 or P＜0.05).The relative quantities of placental MT1 and ZnT1 proteins and mRNA expressions in interventricular septal defect (protein quantities 0.438±0.096 and 0.384±0.061,mRNA expressions 1.23±0.82 and 0.96±0.39),atrial septal defect (0.427±0.093 and 0.377±0.059,1.17±0.70 and 0.85±0.40) and tetralogy of Fallot (0.414±0.111 and 0.336±0.066,1.31±0.97 and 0.90±0.38) groups were significantly lower than those in their corresponding control groups (protein quantities 0.565±0.083 and 0.541±0.090,mRNA expressions 2.78± 1.06 and 1.67±0.33;protein quantities 0.622±0.136 and 0.493±0.079,mRNA expressions 2.85±0.89 and 1.72±0.38;protein quantities 0.637±0.125 and 0.521±0.089,mRNA expressions 3.21 ± 0.99 and 1.61±0.29;P＜0.01 or P＜0.05).Conclusion Mothers with their fetus of certain types of CHD are found zinc deficiency,and down-regulation of MT1 and ZnT1 expressions in the serum and placenta may involve in the pathogenesis of CHD when maternal zinc deficiency.

Objective : To investigate the sex difference of the effects of chronic pregnancy stress on depression-like behavior in offspring adolescent mice and whether the amygdala is involved in mediating depression-like behavior and its possible mechanism． Methods : Male and female of C57BL /6J mice were put in cage together．Pregnant mice were randomly divided into normal control group ( CON group) and chronic pregnancy stress group ( CPS group) ．The day of delivery was recorded as post-natal day(PND0) ．The offspring of different groups were divided into Female group and Male groupaccording to sex，respectively．From PND35，the depressive-like behavior of off- spring was monitored in different groups．Morphological structure of basolateral amygdala (BLA) cone neurone was observed by Golgi-Cox staining，and apoptosis of BLA neurone was detected by TUNEL．Serum corticotrophin-relea- sing hormone ( CRH) was detected by ELISA．The level of protein associated with amygdala mammalian target of rapamycin (mTOR) and phosphorylated mammalian target of rapamycin ［p-mTOR ( Ser2448) ］was detected by Western blot. Results : Depression-like behavior was appeared in different sexual offspring by chronic pregnancy stress，and there was an interaction between chronic pregnancy stress and gender．In the forced swimming test，the immobility time of offspring in the CPS group prominently increased(Female: P＜0. 05，Male: P＜0. 001) ．Interest- ingly，compared with female offspring ，despairing behavior of male offspring was much more clearly observed in CPS group(P＜0. 05) ．Compared with offspring of CON group，the rate of sucrose preference was significantly re- duced in the female offspring of CPS group(P＜0. 05) ，while no obvious difference was observed in the male off- spring．Compared with the CON group，the density of neuronal dendrite branches in the BLA of offspring mice in the CPS group decreased(Female: P＜0. 01，Male : P＜0. 01) and the degree of neuronal apoptosis increased( Fe- male: P＜0. 001，Male : P ＜0. 001) ，the expression level of p-mTOR in amygdala of offspring mice in CPS group significantly decreased(Female: P＜0. 001，Male: P＜0. 001) ．Chronic pregnancy stress increased the serum CRH level of offspring mice(P＜0. 001) ，and the gender had significant influence on serum CRH level，the serum CRH level of female in CPS group was higher than that of male(P＜0. 05) ． Conclusion Chronic pregnancy stress leads to depression-like behavior in offspring adolescent mice，and the depression-like behavior has gender differences. In addition，chronic pregnancy stress leads to dendrite atrophy and apoptosis of BLA neurons in offspring mice，and the mechanism may be that the activation of mTOR in the amygdala of offspring mice is inhibited．CRH may be in- volved in mediating sex differences in depression-like behavior and BLA neuron damage in offspring induced by chronic pregnancy stress.