Objective To investigate the role of glucose-regulated protein 78 (GRP78) in sevoflurane preconditioning-induced inhibition of apoptosis in cardiomyocytes of rats.Methods The cultured cardiomyocytes of Sprague-Dawley rats were randomly divided into 5 groups (n =30 each) using a random number table:control group (group C),hypoxia-reoxgenation (H/R) group,sevoflurane preconditioning group (group S),siRNA-GRP78 group and siRNA control group.H/R was produced by 2 h exposure of cells to 95％ N2-5％ CO2 in an air-tight chamber at 37 ℃,followed by reoxygenation with 95％ O2-5％ CO2 in an air-tight chamber at 37 ℃ for 1 h.In group S,the cells were incubated with 2.5％ sevoflurane for 20 min,followed by 10-min washout before H/R.In siRNA-GRP78 group,the cells were transfected with siRNA-GRP78 100 nmol/L,and 24 h later preconditioning with sevoflurane was performed and H/R was produced.In siRNA group,cells were transfected with siRNA,and the other treatments were similar to those previously described in siRNA-GRP78 group.After treatment in each group,the expression of GRP78 in myocardial cells and cytochrome c in cytoplasm and mitochondria was detected by Western blot.Lactic dehydrogenase (LDH) and creatine kinase (CK) activities in the culture medium of each group were determined by ELISA.The apoptosis in myocardial cells was assessed by flow cytometry.Apoptotic rate was calculated.Intracellular free Ca2+ concentration ([Ca2+]i) was measured with the fluorescent probe Fura-2/ AM.The opening of mPTP was measured by fluorescence spectrophotometry.Results Compared to group C,the expression of GRP78 in myocardial cells and cytochrome c in cytoplasm was significantly up-regulated,LDH and CK activities in the culture medium,apoptotic rate and [Ca2+]i were increased,and the expression of cytochrome c in mitochondria was down-regulated in H/R group.Compared to group H/R,the expression of GRP78 in myocardial cells and cytochrome c in mitochondria was significantly up-regulated,LDH and CK activities in the culture medium,apoptotic rate,[Ca2+] i and opening of mPTP were decreased,and the expression of cytochrome c in cytoplasm was down-regulated in group S,and no significant change was found in the parameters mentioned above in siRNA group.Compared to group S,the expression of GRP78 in myocardial cells and cytochrome c in mitochondria was significantly down-regulated,LDH and CK activities in the culture medium,apoptotic rate,[Ca2+] i and opening of mPTP were increased,and the expression of cytochrome c in cytoplasm was up-regulated in group siRNA-GRP78.Conclusion GRP78 is involved in sevoflurane preconditioning-induced inhibition of apoptosis in cardiomyocytes of rats,and the mechanism is related to maintenance of intracellular Ca2+ stability and inhibition of opening of mPTP.

objective　To study the expression of CD44v3 and CD44v6 and its relationship with lymph node metastasis in non small cell lung cancer (NSCLC). Methods　Specimens (lung tissues) from 52 c ases of NSCLS and 12 normal lung tissue were used to detect the expression of CD 44v3 and CD44v6 by immunohistochemical method (SP method) and flow cytometry, co rrelation was analysed between the expression of CD44v3 or CD44v6 a nd lymph node metastasis of the lung cancer. Results　CD44v3 and CD44v6 were not, or weakly expressed in all normal lung tissues from 12 cases. In contrast, the expression levels of CD44v3 and CD44v6 were obviously higher in lung cancer than that in normal tissue(P<0.05). The expression of CD44v 3 and CD44v6 were much higher in patients with lymph node metastasis than those without lymph node metastasis(P<0.05). Conclusion　① CD44v3 and CD44v6 are expressed with different degree in NSCLC. ②There is a close rel ationship between high expression of CD44v6 and lymph node metastasis, and CD44v 6 may be a co-marker for predicting the potentiality of lymph node metastasis i n lung cancer.

Aged ; Angioplasty, Balloon, Coronary ; Heparin ; adverse effects ; Humans ; Male ; Thrombocytopenia ; chemically induced

Objective To synthesize Wye-125132,an inhibitor of mTOR,and to establish a synthetic route for industrial production.Methods Barbituric acid was used as the raw material to synthesize the intermediate 2, 4, 6-trichrolo-pyrimidine-5-carbaldehyde 2 via Vilsmeier-Haack and chlorination reaction, while intermediate 5 was prepared via 3-step reaction from 1,4-dioxaspiro-[4,5]decan-8-one.The condensation of 2 and 5 was followed by substitution reaction to obtain the key intermediate 7.The side chain 9 was prepared by 3-step reaction from bromine aniline.Then the title product 1 was obtained with the reaction of Suzuki cross-coupling of 7 and 9.The structures of intermediate and target compounds were confirmed by MS and 1 H-NMR.Results and Conclusion Compared with the method reported in the literature,this new synthesis method possesses some advantages, such as ready availablity of raw materials, simple operation, mild reaction conditions and easy disposal of products.The total yield is 13.2%,and the purity of the target compound is 99.77%.

Objective:To investigate the clinical and neuroimaging features of atypical teratoid/rhabdoid tumors (AT/RT) of the central nervous system and the survival prognosis in different locations.Methods:The clinical data of 27 patients with AT/RT confirmed by biopsy or postoperative pathology in Sanbo Brain Hospital of Capital Medical University from October 2012 to September 2020 were collected, including 17 males and 10 females, aged (7.6±10.0) years, ranging from 0.2 to 39.0 years old.The clinical features and the results of the first preoperative imaging examination were retrospectively analyzed.The patients were divided into supratentorial, infratentorial and trans supratentorial and infratentorial groups according to the location.The survival time of the three groups was compared by Kaplan-Meier survival curve.Results:All patients presented with headache, including 12 cases with vomiting.There were 12 cases of supratentorial, 10 cases of infratentorial and 5 cases of supratentorial and infratentorial.There were 20 cases of cystic degeneration, 7 cases of calcification, 6 cases of hemorrhage and 13 cases of peritumoral edema.The median survival time of patients with infratentorial AT/RT was longer than that of patients with supratentorial and transtentorial AT/RT (χ 2=7.353, P=0.025). Conclusion:Central nervous system AT/RT is easy to occur in young children, and the survival time of AT/RT patients is longer.

Objective:To investigate the anti-oxidative stress effects of microRNA 125b (miR-125b) on lens epithelial cells (LECs) and its possible mechanism.Methods:Twenty-four anterior capsule specimens were collected from 24 eyes of 24 age-related cataract patients during phacoemulsification and 20 normal anterior capsule specimens were obtained from 20 eyes of 20 donors in Henan Eye Hospital from July 2018 to March 2019 under the approval of a Medical Ethics Committee of Henan Eye Hospital (No.YKYY20193151).The reverse transcription PCR and Western blot assay were employed to detect and compare the relative expression levels of miR-125b and nuclear factor E2-related factor 2 (Nrf2) in different specimens.The human lens epithelial cell line HLEB-3 was divided into control group and oxidative stress model group.The oxidative stress models were established by coculture with different concentrations (100, 200, 400 μmol/L) of H 2O 2 for 24 hours, and the cells were cultured with normal medium without H 2O 2 in the control group.The reactive oxygen species (ROS) content was detected by DCFH-DA fluorescent probe, and the activities of total-antioxidative capability (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) as well as malondialdehyde (MDA) concentration were detected by ELISA, and compared among the groups.The expression levels of miR-125b and Nrf2 were detected by reverse transcription PCR and Western blot assay, respectively.The cells were transfected with miR-125b mimics, miR-125b control and miR-125b inhibitor for 24 hours, respectively, and ROS content was detected by DCFH-DA fluorescent probe and T-AOC, SOD and GSH-Px activities as well as MDA concentration were detected by ELISA and compared among different transfected groups.A dual luciferase reporter assay was used to assess an association between miR-125b and Nrf2.The expression level of Nrf2 protein was detected by Western blot assay and the expression levels of Nrf2 and Keap1 were assayed and located by immunofluorescence double staining. Results:The relative expression levels of miR-125b and Nrf2 in the normal lens anterior capsule specimens were 0.21±0.03 and 0.27±0.06, which were significantly lower than 0.89±0.05 and 0.84±0.12 in the cataract specimens, respectively ( t=15.355, P<0.05; t=18.647, P<0.05).The relative expression levels of miR-125b and Nrf2 were significantly increased in various H 2O 2 treated groups in comparison with the control group and were gradually elevated with the increase of H 2O 2 concentration (all at P<0.05).Compared with the control group, the T-AOC, SOD and GSH-Px activities were reduced, and ROS content and MDA concentration were significantly ascended (all at P<0.05).Compared with the miR-125b control group, the T-AOC, GSH-Px and SOD activities were increased, and ROS content and MDA concentration were decreased in the miR-125b mimics group (all at P<0.05).In addition, the T-AOC, GSH-Px and SOD activities were significantly weakened, and ROS content and MDA concentration were significantly increased in the miR-125b inhibitor group in comparison with the miR-125b control group (all at P<0.05).Dual luciferase reporter assay showed that miR-125b targeted to the expression of Nrf2 in the H 2O 2 model cells.The fluorescence of Nrf2 in the cytoplasm was the strongest with more nuclear transfer in the miR-125b mimics group, and the expression intensity of Keap1 in the cytoplasm was weaker.The expression of Nrf2 was the weakest with less nuclear transfer in the miR-125b inhibitor group, and the expression level of Keap1 in the cytoplasm was stronger. Conclusions:MiR-125b can enhance the anti-oxidative stress of LECs in age-related cataractous eyes probably by upregulating the expression of Nrf2 and activating the Keap1/Nrf2 signaling pathway.

ObjectiveTo observe the effects of XinShu parenteral solution (XSPS) on the activation of platelet, the activity of fibrinolysis system after intima denudation of rabbits.Methods20 male Japanese white rabbits (2.5±0.5)kg were randomly divided into the control group and XSPS group .The celiac arterial endothelium of all rabbits were denuded with balloon. Before the operation and 3d, 7d, after balloon denudation, vein blood samples were obtained from each group rabbits for measurement of α granule membrane protein of platelets(GMP-140), tissue-type plasminogen activator(t-PA) and plasminogen activator inhibitor-type Ⅰ(PAI-1).ResultsPlasma GMP-140 and PAI-1 activity obviously elevated after balloon injury, and there was a little elevation in plasma t-PA activity in control group. Activity of plasma GMP-140 in XSPS group remained bottom level after balloon injury, and there was a significant increase in plasma t-PA activity and a marked reduce in PAI-1 activity in XSPS group. There was a notable difference between group B and group C (P<0.05). ConclusionXSPS obviously inhibits platelet activation, and improves fibrinolysis activity after balloon injury.

Objective To investigate the relationship between bed rest time and development of prethrombotic state in the elderly patients with hip fracture.Methods One hundred and sixty-six patients who stayed in bed after hip fracture,aged ≥65 yr,were divided into 5 groups according to the bed rest time on admission to hospital:bed rest time ＜ 24 h (Ⅰ group,n =61),bed rest time 24-48 h (Ⅱ group,n =29),bed rest time 3-6days (Ⅲ group,n =29),bed rest time 7-14 days (Ⅳ group,n =34),and bed rest time ＞ 14 days (Ⅴ group,n =13).Venous blood samples were collected to measure the platelet count,coagulation function,and concentrations of plasma D-Dimer and serum α-granule membrane protein 140 (GMP-140).The development of abnormality in each index was recorded.The development of deep vein thrombosis in both lower extremities was diagnosed using color Doppler ultrasound in D-Dimer-positive patients.Results Compared with group Ⅰ,the abnormal rate of fibrinogen (Fib) and D-Dimer and serum GMP-140 concentrations were significantly increased in Ⅱ and Ⅲ groups,the abnormal rate of platelet count,Fib and D-Dimer and serum GMP-140 concentrations were increased in lⅣ group,and the abnormal rate of platelet count was increased,and no significant change was found in the serum GMP-140 concentrations and abnormal rate of Fib and D-Dimer in Ⅴ group.Compared with group Ⅱ,the serum GMP-140 concentrations were significantly increased in Ⅲ and Ⅳ groups,the abnormal rate of Fib and D-Dimer was increased in Ⅳ group,and no significant change was found in the abnormal rate of Fib and DDimer in Ⅲ group.The abnormal rate of platelet count was significantly lower in Ⅳ group than in Ⅲ group.Color Doppler Ultrasonography showed no sign of deep vein thrombosis.Conclusion For the elderly patients with hip fracture,the possibility of pre-thrombotic state developed is increased when the bed rest time is more than 24 h,and the patients were classified as high-risk patients when the bed rest time is more than 3 days.

AIM:ToinvestigatetheexpressionofFoxp3+regulatoryTcells(Foxp3+Tregs)andprogrammed death receptor 1 (PD1) in gastric cancer tissues and their association with clinicopathological factors and prognosis of the patients.The correlation between the 2 molecules was also analyzed at the same time.METHODS: The tumor sections from 111 gastric cancer patients were stained for Foxp3 and PD1 by the method of immunohistochemistry.The associations of the expression levels of these 2 molecules with clinicopathological factors involved in the disease progression and progno-sis were statistically analyzed .The relationship of their expression was detected.RESULTS:Foxp3 +Tregs and PD1 were expressed in the gastric cancer tissues, and PD1 was expressed in the tumor infiltrating lymphocytes ( TILs) .The expres-sion of Foxp3 and PD1 was correlated with lymph node metastasis, clinicopathological stage and prognosis of gastric cancer patients.The expression of these 2 determinants in the patients with lymph node metastasis and an advanced clinicopatho-logical stage was distinctly higher ( P<0.05 ) .The patients with positive expression of the 2 indexes presented a lower overall survival rate and worse prognosis (P<0.05).A significantly positive correlation between the infiltration of Foxp3 +Tregs and the expression of PD1+TILs was also observed (P<0.01).CONCLUSION:Foxp3 +Tregs and PD1 +TILs co-infiltrate in the gastric cancer tissues, which can be used as biological markers to predict the disease progression and prog-nosis.

Objective To investigate the expression of activated ERK (p-ERK) and activated p38 (p-p38) in the keratinocytes of condyloma acuminata (CA) lesions. Methods Fifty cases of HPV 6/11 CA were diagnosed by in situ hybridization. The expression and distribution of p-ERK and p-p38 in CA lesions and 25 normal human skins (foreskins) were detected by immunohistochemistry technique (En Vision). Results ①The results showed that the expression of p-ERK and p-p38 in keratinocytes of CA lesions were significantly higher than those in normal epidermis (P