1.Effect of Compatibility of Effective Monomer Components of Fujin Shengjisan on Angiogenesis of HUVEC Based on Uniform Design
Xianying LU ; Jing GAO ; Dingxi BAI ; Chaoming HOU ; Wenting JI ; Huan CHEN ; Chenxi WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):9-20
ObjectiveTo determine the optimal combination of the effective monomer components "quercetin-kaempferol-abietic acid-boswellic acid" in Fujin Shengjisan for promoting diabetic ulcer (DU) wound healing through uniform design, thereby achieving the modern application of the ancient formula. MethodsFollowing the principle of "uniform design-pharmacodynamic experiment-mathematical modeling and model verification", the U14(145) uniform design table was adopted.The four monomer components of Chinese medicine were considered as the independent variables, and the proliferation rate of human umbilical vein endothelial cells (HUVECs) induced by glucose was used as the pharmacodynamic indicator. A mathematical model was constructed using DPS software to correlate the effective monomer components with the pharmacodynamic indicator. The results of uniform design were verified through CCK-8 assay, cell scratch healing, tube formation, Western blot, and Real-time PCR. ResultsAmong the 14 compatibility groups, compared with the high-glucose model group, compound compatibility group 6 showed the strongest proliferation effect and statistical significance (P<0.05). Four quadratic polynomial regression equations (Y1-Y4) were obtained through DPS modeling. Considering the model's fit, stability, and practical application, equations Y1-Y3 were selected for the follow-up verification. To ensure experiment reproducibility, group 6 was used for validation. Group 6 and equations Y1-Y3 were renamed as compound prescription ① to compound prescription④, respectively, to represent the modern application of the ancient FJSJ Powder through compatibility of monomer components. Verification experiments showed that in the CCK-8, scratch healing, and tube formation assays, the cell viability, wound healing rate, and tube formation number of HUVECs stimulated with 50 mmol·L-1 glucose were significantly reduced compared with the blank group. Moreover, the expression levels of angiogenesis-related cytokines, vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2), and CD31 secretion were significantly down-regulated. However, after intervention with compound prescriptions ① to ④, compound prescriptions ① and ③ significantly improved the biological functions of HUVECs induced by 50 mmol·L-1 glucose. Further analysis of the regression coefficients of compound prescriptions ① and ③, and the relative dose ratios of each monomer component, indicated that abietic acid, quercetin, and boswellic acid promoted angiogenesis of HUVECs in the high glucose environment, with a major effect (positive partial correlation coefficients, all > 0.9). Abietic acid and boswellic acid, as well as kaempferol and boswellic acid, promoted angiogenesis in HUVECs through interaction (positive partial correlation coefficients). ConclusionCompound prescriptions ① and ③ are the optimal combinations. They can reverse the inhibitory effects of high glucose, stimulate the proliferation, migration, and tube formation abilities of HUVECs in a high glucose environment, and promote the expression of vascular endothelial growth factorA(VEGFA), FGF2, and CD31, thereby promoting angiogenesis and facilitating DU wound healing. This finding not only confirms the good reproducibility and feasibility of compound prescriptions ① and ③ but also provides new insights and methods for the rational construction of mathematical models to further study the compatibility theory of Chinese medicine.
2.Efficacy evaluation of denosumab in the treatment of patients with postmenopausal osteoporosis at high risk of fracture
Jing ZHOU ; Xingyun HOU ; Tingting LIU ; Jiaoyang ZHENG
Chinese Journal of Clinical Medicine 2025;32(1):85-92
Objective To explore the difference of efficacy and safety between denosumab and zoledronic acid in the treatment of patients with postmenopausal osteoporosis (PMOP), and to optimize the medication regimen for PMOP patients. Methods A total of 123 PMOP patients with high risk of fracture at the Second Affiliated Hospital of Naval Medical University from September 2021 to March 2024 were selected and randomly divided into two groups: the denosumab group (n=63) and the zoledronic acid group (n=60). Both groups underwent one-year treatment and follow-up, bone metabolism indexes, lumbar vertebrae, femoral neck, and total hip bone mineral density (BMD) were monitored, and any adverse reactions were documented. Results After treatment, the lumbar vertebrae and total hip BMD of patients in the denosumab group and the zoledronic acid group were significantly improved (P<0.05); the femoral neck BMD of patients in the zoledronic acid group was also significantly improved (P<0.05). The improvement of lumbar vertebrae BMD in the denosumab group was significantly better than that in the zoledronic acid group, while the improvement of femoral neck and total hip BMD in the zoledronic acid group was significantly better than that in the denosumab group (P<0.05). Bone metabolism indicators were significantly improved in both groups (P<0.05), and no significant liver and kidney dysfunction were observed. A total of 7 patients in the zoledronic acid group had mild adverse reactions and 5 patients in the denosumab group had mild adverse reactions. Conclusions Denosumab significantly increased lumbar vertebrae BMD and improved bone metabolism markers in PMOP patients, thus reducing risk of fracture and demonstrating good safety.
3.Construction of PD-L1hitol-DC derived from bone marrow of DA rats and identification of its immunological function
Zhiqi YANG ; Peibo HOU ; Lang WU ; Jing LIU ; Yang DING ; Minghao LI
Organ Transplantation 2025;16(1):83-90
Objective To construct programmed cell death protein-ligand 1(PD-LI)hi tolerogenic dendritic cell (tol-DC) derived from bone marrow of DA rats and identify its immunological function. Methods DA rat bone marrow cells were extracted, combined with recombinant mouse granulocyte macrophage colony-stimulating factor and recombinant mouse interleukin (IL)-4, and cultured for 6 days in vitro to induce the differentiation of bone marrow cells into immature dendritic cells (imDC). Lipopolysaccharide was used to stimulate cell maturation and cultured for 2 days to collect mature dendritic cells (mDC). PD-L1 lentiviral vector virus stock solution or equivalent dose lentiviral stock solution was added, and PD-L1hitol-DC and Lv-imDC were collected after culture for 2 days. The morphology of PD-L1hitol-DC was observed by inverted phase contrast microscope and transmission electron microscope. Real-time fluorescence quantitative reverse transcription polymerase chain reaction, Western blotting and flow cytometry were used to detect the expression level of specific markers on cell surface. CD8+T cells derived from Lewis rat spleen were co-cultured with imDC, mDC, Lv-imDC and PD-L1hitol-DC, respectively. The levels of inflammatory factors in the supernatant of each group were detected by enzyme-linked immunosorbent assay. The apoptosis of T cells and the differentiation of regulatory T cells (Treg) in each group were analyzed by flow cytometry. Results The morphology of PD-L1hitol-DC modified by PD-L1 gene was consistent with tol-DC characteristics, and the expression levels of CD80, CD86 and major histocompatibility complex (MHC) on the surface were low. After mixed culture with CD8+ T cells, the levels of IL-10 and transforming growth factor (TGF) -β1 in the supernatant of PD-L1hitol-DC group were higher, the levels of tumor necrosis factor (TNF) -α and IL-17A were lower, and the apoptosis of T cells and Treg differentiation were increased. Conclusions Overexpression of PD-L1 through lentiviral vectors may successfully induce the construction of bone-marrow derived PD-L1hitol-DC in DA rats, promote the secretion of anti-inflammatory factors and T cell apoptosis, induce the differentiation of Treg, and inhibit the immune response of allogeneic CD8+T cells, which provides experimental basis for the next organ transplantation immune tolerance study.
4.Effect of Spraying Nano-calcium Carbonate and Calcium Nitrate Tetrahydrate on Characters, Carbohydrate Components and Endogenous Hormones of Dendrobium officinale
Jing LI ; Rong ZHOU ; Yingyue HOU ; Wei CAI ; Xiaoyan ZHANG ; Shuang ZHANG ; Guangying DU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):208-216
ObjectiveTo investigate the effects of foliar fertilizer of nano-calcium carbonate and calcium nitrate tetrahydrate on the agronomic traits, carbohydrate and endogenous hormone contents of Dendrobium officinale planted for 1 year under greenhouse cultivation, in order to provide scientific basis for fertilization to improve the yield and quality of D. officinale. MethodsSingle-factor experimental design was adopted. Starting from early spring, D. officinale was treated with foliar spraying according to corresponding fertilizers. Three treatment groups were established based on different fertilizers, namely, a blank group(clear water), a nano-calcium carbonate group(0.727 g·L-1 nano-calcium carbonate water-soluble fertilizer), and a calcium nitrate tetrahydrate group(1.091 g·L-1 calcium nitrate tetrahydrate water-soluble fertilizer). The frequency of spraying was three times per month, and the entire treatment process lasted for nine months. The effects of various treatments on the traits and relative chlorophyll content of D. officinale were dynamically monitored. Sampling was conducted at three specific time points:August 2, 2023, September 8, 2023, and November 1, 2023, respectively. The contents of glucose and mannose in D. officinale stems were determined by high performance liquid chromatography(HPLC), the content of soluble sugars in D. officinale stems and leaves was determined by phenol method, and enzyme-linked immunosorbent assay(ELISA) was used to detect the concentrations of cytokinin and auxin. ResultsCompared with the blank group, the treatments with nano-calcium carbonate and calcium nitrate tetrahydrate could significantly increase stem length, stem node number, leaf number, and tiller number. Among them, during the harvesting period in November, the stem length and tiller number, which are indicators related to the yield of D. officinale, increased by 60.85% and 19.23% after treatment with calcium nitrate tetrahydrate, and by 32.54% and 28.85% after treatment with nano-calcium carbonate, respectively. Compared with the blank group, treatments with nano-calcium carbonate and calcium nitrate tetrahydrate could promote the accumulation of sucrose in the stems and leaves of D. officinale to varying degrees, as well as the accumulation of polysaccharides, mannose, and glucose in the stems. In addition, nano-calcium carbonate treatment also facilitated the accumulation of fructose in the stems and leaves of D. officinale. Specifically, during the harvesting period in November, polysaccharides and mannose, which were the main active ingredients in D. officinale stems, increased by 28.48% and 29.36% after treatment with calcium nitrate tetrahydrate, and by 39.91% and 82.62% after treatment with nano-calcium carbonate, respectively. In addition, compared with the blank group, the concentrations of auxin in the stems and leaves of D. officinale were significantly increased after treatment with calcium nitrate tetrahydrate(P<0.05). Similarly, the concentrations of cytokinin and auxin in the stems of D. officinale were also elevated after treatment with nano-calcium carbonate. Correlation analysis further indicated that elongation growth and tillering of D. officinale stems after foliar spraying of nano-calcium carbonate and calcium nitrate tetrahydrate might be related to the accumulation of carbohydrates in the stems and leaves and the synergistic effect of auxin and cytokinin. ConclusionIn production practice, spraying nano-calcium carbonate and calcium nitrate tetrahydrate can promote the accumulation of cytokinin, auxin, and carbohydrate contents in the stems and leaves of D. officinale, and promote tillering and elongation growth of the stems.
5.Research progress of Faricimab in the treatment of macular edema associated with retinal vascular diseases
Xinyi HOU ; Haoran WANG ; Chunhua DAI ; Jing ZHANG ; Meng XIN ; Zhixin GUAN ; Shu LIU
International Eye Science 2025;25(8):1267-1273
Intravitreal injection of anti-vascular endothelial growth factor(VEGF)agents has become the primary treatment for macular edema associated with retinal vascular disease such as diabetic retinopathy and retinal vein occlusion, but there are limitations such as variable treatment efficacy and insufficient durability of therapeutic effects. As the first bispecific antibody applied in ophthalmic treatment, Faricimab achieves favorable outcomes by simultaneously targeting both VEGF-A and angiopoietin-2(Ang-2)pathways. Based on evidence from recent clinical trials and real-world studies, this article reviews the research progress on Faricimab for the treatment of diabetic macular edema(DME), retinal vein occlusion-associated macular edema(RVO-ME)and refractory macular edema compared to the therapeutic effects of other agents. Additionally, based on Faricimab's safety characteristics and future potential, its therapeutic prospects for macular edema associated with retinal vascular diseases are discussed. This review aims to provide evidence-based references for optimizing clinical treatment strategies, thereby contributing to mitigating the risk of vision loss due to macular edema.
6.Allogeneic intrastromal lenticule implantation combined with corneal collagen cross-linking for moderate to advanced keratoconus
Jing ZHANG ; Jie HOU ; Yahui DONG ; Yulin LEI ; Yafei XU ; Fangfang SUN
International Eye Science 2025;25(9):1517-1522
AIM: To evaluate the safety and efficacy of allogeneic intrastromal lenticule implantation combined with corneal collagen cross-linking(CXL)in patients with moderate to advanced keratoconus.METHODS: A retrospective case series analysis was conducted. A total of 19 patients(20 eyes)with moderate to advanced keratoconus who underwent combined allogeneic intrastromal lenticule implantation and CXL at the Jinan Mingshui Eye Hospital from June 2021 to December 2023 were included. The uncorrected distance visual acuity(UCVA), thinnest corneal thickness, central corneal epithelial thickness, anterior corneal flat keratometry(Kf), steep keratometry(Ks), and mean keratometry(Km), as well as the first applanation time(A1T), the first applanation length(A1L), the velocity during the first applanation moment(VIN), the second applanation time(A2T), the second applanation length(A2L), the velocity during the second applanation moment(VOUT), highest concavity time(HCT), highest concavity radius(HCR), peak distance(PD), deformation amplitude(DA), stiffness parameter at first applanation(SP-A1), integrated radius(IR), central corneal thickness(CCT), intraocular pressure(IOP), corneal thickness-corrected IOP, biomechanically intraocular pressure IOP(bIOP), and corneal thickness variation rate(ARTH)were compared between the two groups before surgery and at 1 wk, 1, 3 and 6 mo after surgery.RESULTS: All patients successfully completed the surgery without any intraoperative complications. No significant differences were observed between pre-operative and post-operative measurements for UCVA or the corneal biomechanical parameters, including A1L, A2L, PD, A1T, A2T, VIN, VOUT, DA, IOP, and bIOP(all P>0.05). Significant differences were found between pre-operative and post-operative values for corneal thinnest point thickness, central corneal epithelial thickness, Kf, Ks, Km, and the corneal biomechanical parameters, including HCT, HCR, SP-A1, ARTH, IR, and CCT(all P<0.05). The anterior corneal curvature demonstrated an initial increase followed by a decrease post-operatively. Furthermore, significant differences were observed between pre-operative and post-operative values for HCT, HCR, SP-A1, ARTH, IR, and CCT(all P<0.005).CONCLUSION: Allogenic intrastromal lenticule implantation combined with corneal collagen cross-linking demonstrates favorable safety and stability in treating moderate-to-advanced keratoconus. This combined procedure effectively increases corneal thickness and rigidity, resulting in corneas that are more resistant to deformation postoperatively.
7.Assessment of cheese sign and its association with vascular risk factors: Data from PUMCH dementia cohort
Xinying HUANG ; Bo HOU ; Jie WANG ; Jie LI ; Li SHANG ; Chenhui MAO ; Liling DONG ; Caiyan LIU ; Feng FENG ; Jing GAO ; Bin PENG
Chinese Medical Journal 2024;137(7):830-836
Background::In the clinic, practitioners encounter many patients with an abnormal pattern of dense punctate magnetic resonance imaging (MRI) signal in the basal ganglia, a phenomenon known as "cheese sign". This sign is reported as common in cerebrovascular diseases, dementia, and old age. Recently, cheese sign has been speculated to consist of dense perivascular space (PVS). This study aimed to assess the lesion types of cheese sign and analyze the correlation between this sign and vascular disease risk factors.Methods::A total of 812 patients from Peking Union Medical College Hospital (PUMCH) dementia cohort were enrolled. We analyzed the relationship between cheese sign and vascular risk. For assessing cheese sign and defining its degree, the abnormal punctate signals were classified into basal ganglia hyperintensity (BGH), PVS, lacunae/infarctions and microbleeds, and counted separately. Each type of lesion was rated on a four-level scale, and then the sum was calculated; this total was defined as the cheese sign score. Fazekas and Age-Related White Matter Changes (ARWMC) scores were used to evaluate the paraventricular, deep, and subcortical gray/white matter hyperintensities.Results::A total of 118 patients (14.5%) in this dementia cohort were found to have cheese sign. Age (odds ratio [OR]: 1.090, 95% confidence interval [CI]: 1.064-1.120, P <0.001), hypertension (OR: 1.828, 95% CI: 1.123-2.983, P = 0.014), and stroke (OR: 1.901, 95% CI: 1.092-3.259, P = 0.025) were risk factors for cheese sign. There was no significant relationship between diabetes, hyperlipidemia, and cheese sign. The main components of cheese sign were BGH, PVS, and lacunae/infarction. The proportion of PVS increased with cheese sign severity. Conclusions::The risk factors for cheese sign were hypertension, age, and stroke. Cheese sign consists of BGH, PVS, and lacunae/infarction.
8.A multicenter clinical study on the treatment of lateral epicondylitis of humerus by manipulation
Xiao-Zhou HOU ; Jing YIN ; Hai-Yang WANG ; Jin-Yu GU ; Tian-Hao WAN ; Man-Hong YANG ; Di XIA ; Qing ZHANG
China Journal of Orthopaedics and Traumatology 2024;37(3):251-257
Objective To investigate clinical effect of tendons pulling,poking and kneading for the treatment of external humeral epicondylitis.Metods From January 2018 to December 2021,a multicenter randomized controlled study was per-formed to collect 192 patients with external humeral epicondylitis in Wangjing Hospital,Beijing Dianli Hospital,and Beijing Fengsheng Osteotraumatology Hospital,respectively,and they were divided into treatment group and control group by random number table method.There were 96 patients in treatment group,including 36 males and 60 females,aged from 28 to 60 years old with an average of(41.20±5.50)years old;the course of disease ranged from 1 to 14 days with an average of(5.24±1.35)days;they were treated once every other day for 2 weeks.There were 96 patients in control group,including 33 males and 63 females,aged from 26 to 60 years old with an average of(43.35±7.75)years old;the course of disease ranged from 1 to 14 days with an average of(5.86±1.48)days;they were treated with topical voltaalin combined with elbow joint fixation for 2 weeks.Visual analogue scale(VAS)and Hospital for Surgery Scoring System(HSS)elbow pronation and supination angles,wrist metacarpal flexion and dorsal extension angles,elbow tenderness between two groups were compared before treatment and at 1,3,5,7,11 and 13 days after treatment;Hospital for Surgery Scoring System 2(HSS2)was compared before treatment and the final treatment.Results All patients were followed up for 10 to 14 days with an average of(12±1.6)days.VAS between treatment group and control group before treatment were 6.83±1.36 and 6.79±1.58,respectively,and decreased to 1.49±1.09 and 2.11±1.81 after the final treatment.VAS of treatment group were significantly lower than those of control group at 1,3,5,7,9,11 and 13 days after treatment(P<0.05).HSS between two groups were 61.73±11.00 and 36.47±12.45 before treatment,respectively,and increased to 94.42±5.9 and 91.44±9.11 at the final treatment.HSS of treatment group were signifi-cantly higher than those of control group at 1,3,5,7,9,11 and 13 days after treatment(P<0.05).On the 5th day after treat-ment,the external and internal rotation angles of elbow in treatment group were(66.41±12.69)° and(66.35±13.54)°,while those in control group were(62.08±16.03)° and(61.77±16.35)°.On the 7th day after treatment,the external and internal ro-tation angles of elbow were(69.79±12.64)° and(70.02±13.55)° in treatment group,and(65.28±15.86)° and(65.09± 16.67)° in control group.Elbow joint motion in treatment group was higher than that in control group(P<0.05).On the 5th day after treatment,angles of wrist dorsiflexion and palm flexion were(39.43±15.94)°and(46.68±11.10)° in treatment group,and(38.51±18.49)° and(44.27±13.58)° in control group.On the 7th day after treatment,angles of wrist dorsiflexion and palm flexion were(42.52±16.50)° and(49.23±10.96)° in treatment group,and(41.18±20.09)° and(46.64±14.63)° in control group.The motion of wrist joint in treatment group was higher than that in control group(P<0.05).On the 13th day after treatment,HSS2 in treatment group 93.61±6.32 were higher than those in control group 92.06±7.94(P<0.05).There was no significant difference in elbow tenderness between two groups at each time point(P>0.05).Conclusion Voltaren external treatment combined with elbow fixation and tendons pulling,poking and kneading could effectively improve symptoms of exter-nal humeral epicondylitis.Compared with voltaren external treatment,tendons pulling,poking and kneading has advantages of longer analgesic time and better elbow function recovery.
9.Progress on treatment of acetabular quadrilateral plate injury
Jing-Wei XIAO ; Ming LI ; Dian WANG ; Xiao-Dong GUO ; Zhi-Yong HOU
China Journal of Orthopaedics and Traumatology 2024;37(3):319-326
Acetabular quadrilateral plate injury has become a hot spot and focus in the field of orthopaedic trauma and pelvic floor function in recent years.Although there are five fracture types,they are all based on fracture morphology,without considering the pulling force of ligaments,joint capsular and muscles.A perfect classification needs to describe the displace-ment of bone mass in three-dimensional space to better guide reduction and fixation.The seven incision and exposure methods are still the traditional open-eye surgery,and how to protect the criss-crossing vascular neural network and pelvic organs is still the focus.Quadrilateral defect causes dislocation of artificial hip joint,and quantitative evaluation of quadrilateral defect vol-ume and revision techniques are still a hot topic.In this paper,the viewpoints of three-dimensional network structure of acetab-ular pelvic vascular anatomy,anatomical surgical target channel and fixation anchor point of acetabular fracture reduction are proposed to design new techniques for accurate and minimally invasive surgical operations,in order to realize the requirements of rapid orthopedic rehabilitation.
10.The role of lncRNA MALAT1 and NEAT1 in neuroprotection of hypoxia preconditioning mouse hippocampus cell
Haidong HOU ; Lei YAN ; Liping WANG ; Jing YANG ; Yucheng GUI ; Yongqiang DU ; Guo SHAO
Chinese Journal of Cerebrovascular Diseases 2024;21(8):525-536
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.

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