Objective The present study was designed to explore the inhibitory effect of antisense VEGF-(165) cDNA on angiogenesis,growth rate of human neuroblastoma.Methods The eukaryotic expression vectors bearing antisense VEGF-(165) cDNA or sense VEGF-(165) cDNA were constructed.The stable cell lines transfected with the sense or antisense VEGF-(165) cDNA were established by using the selective medium containing 400?mg/L of G418.These cell lines were further studied for the exogenous antisense VEGF mRNA expression by RT-PCR and inhibition of expression of endogenous VEGF protein by immunocytochemical staining and ELISA.The proliferation of the transfected cells was analyzed by MTT method.Transfected cells were subcutaneously transplanted into nude mice and the growth of tumor masses was observed and weighted.Results The antisense VEGF was only detected in SH-SY5Y/AsVEGF cells by RT-PCR.The expression of VEGF protein was dramatically declined in the SH-SY5Y/AsVEGF cells compared with the original cells and empty vector transfected cells by immunocytochemical staining and ELISA.There was no effect of antisense VEGF transfection on cell proliferation in vitro.The growth of tumor mass with the cells transfected with antisense VEGF was significantly decreased in nude mice.Conclusion Antisense VEGF cDNA transfection to SH-SY5Y cells can effectively inhibit the expression of endogenous VEGF protein and tumor growth in nude mice.

Objective: To explore early diagnostic value of combined detection of ischemia-modified albumin and other biochemical markers for acute coronary syndrome (ACS). Methods: A total of 156 patients, who received treatment within 4h because of chest pain onset, were divided into ACS group (n=112) and non-ischemic chest pain group (NICP group, n=44) according to coronary angiography (CAG). Blood sample was taken within 4h and during 4~8h after chest pain to measure cardiac biochemical markers, including ischemia-modified albumin (IMA), LDH, CK, CK-MB and cardiac troponin I (cTnI), comprehensive analysis was performed by detected results of above biochemical markers and CAG. Results: For diagnosis of ACS, within 4h after onset IMA possessed highest sensitivity (87.50%) and highest accuracy (80.77%), cTnI possessed highest specificity (95.45%); In 4～8h IMA still possessed highest sensitivity (91.07%), cTnI still possessed highest specificity（97.73%）, CK（85.90%），and IMA（85.25%）possessed highest accuracy （both CK and IMA had no significant difference）; According to comprehensive analysis, regardless of within 4h or during 4~8h after chest pain, IMA possessed highest sensitivity and highest accuracy, cTnI possessed highest specificity. Within 4h after chest pain, detection of IMA combined above-mentioned other four indexes increased sensitivity (89.28%), specificity （95.45%） and accuracy （91.02%）to highest level. Conclusion: For diagnosis of ACS, IMA possesses highest sensitivity and highest accuracy, cTnI possesses highest specificity; IMA combined above-mentioned other four indexes may increase sensitivity, specificity and accuracy to highest level.

Objective To investigate the inhibitive effect of antisense VEGF165 cDNA on growth and angiogensis of human neuroblastoma in nude mouse models.Methods Three kinds of SH-SY5Y cells which had been stably transfected by sense VEGF165 cDNA,antisense VEGF165 cDNA and empty vector respectively,were inoculated subcutaneously to nude mice.The size of subcutaneous tumors was measured,and the morphology of tumor tissue was observed by microscope.The microvessel density(MVD)in tumor mass was analyzed by CD31 immunohistochemistry staining.Apoptotic cells were detected by the electron microscope.Results The growth of tumor mass,which was inoculated with antisense VEGF,significantly decreased in nude mice compared with empty vector group(P

Objective:To evaluate the effect of sevoflurane on necroptosis in isolated hippocampal neurons and the relationship with ryanodine receptor.Methods:Primarily cultured hippocampal neurons of fetal rats of Sprague-Dawley rats were inoculated in culture wells (100 μl/well) or culture flasks (3 ml/bottle) at a density of 5×10 5 cells/ml at 7 days of culture and divided into 3 groups ( n=24 each) using a random number table method: control group (group C), sevoflurane group (group S) and ryanodine receptor antagonist group (group R). Group C received routine culture.Ryanodine receptor antagonist Dantrolene at a final concentration of 3 μmol/L was added in group R. Thirty minutes later, the cells were placed in the incubator containing 2% sevoflurane and cultured for 5 h at 37 ℃ in S and R groups.Then cells were collected, the morphology of neurons was observed with an inverted microscope, the concentrations of free calcium ion ([Ca 2+ ] i) in cytoplasm were determined by flow cytometry, the expression of ryanodine receptor and phosphorylated MLKL protein (p-MLKL) was detected by Western blot, the expression of RIP3 was measured by immunofluorescence, and necroptosis rate was calculated. Results:Compared with group C, the [Ca 2+ ] i were significantly increased, the expression of ryanodine receptor and p-MLKL was up-regulated, and the necroptosis rate was increased in S and R groups ( P<0.05). Compared with group S, the expression of ryanodine receptor and p-MLKL was significantly down-regulated, and the [Ca 2+ ] i and necroptosis rate were decreased in group R ( P<0.05). There was no obvious abnormality in the morphology of neurons in group C. The cell body of neurons were shrunk, the processes were broken, and the network between processes was sparse in group S. The cell body was round, and the morphology was close to normal in group R. Conclusion:Sevoflurane can cause necroptosis in isolated hippocampal neurons of rats, and the mechanism is related to up-regulating the expression of ryanodine receptors and leading to calcium overload.

Objective To study the roles of micheliolide on ovarian cancer cells. Methods Firstly, human ovarian cancer cell lines HeyA8, SKOV3 and A2780/DDP were treated with different concentration of micheliolide (0.25, 0.5, 1, 2.5, 5, 10, 20, 50 μmol/L) for 72 hours, then methyl thiazolyl tetrazolium (MTT) assay was used wo detect the growth of the human ovarian cancer cell lines and the stongest inhibited cell line were selected for the following test. Secondly, after HeyA8 cell line was treated with different concentration (5, 10, 20μmol/L) of micheliolide for 24 hours, the HeyA8 cell apoptosis was measured byflow cytometry. Thirdly, the expression of RelA mRNA in HeyA8 cell was detected through real-time PCR, the expressions of nuclear factor κB(NF-κB)signal pathway related protein RelA and the activited cysteinyl aspartate specific proteinase (caspase-9) were detected by western blot analysis. Results (1) The growth of HeyA8, SKOV3 and A2780/DDP cells were all significantly inhibited after being treated with different concentration of micheliolide for 72 hours and the roles of inhibition were all concentration dependant (P<0.05). The half maximal inhibitory concentration (IC50) of HeyA8, SKOV3 and A2780/DDP were (9.8±2.2), (12.0±2.1) and (12.8±1.8)μmol/L, respectively. We chose HeyA8 cell to do the following expreriments because of its best inhibited effect. (2) After HeyA8 cell was treated with micheliolide of different concentrations, as the concentration increased (20 and 0μmol/L, for example), the apoptosis rate of HeyA8 cell raised from (7.2±1.0)%to (17.4±1.1)%, the percentage of survived cells reduce from (92.8 ± 1.3)% to (82.6 ± 1.4)%,and the relative mRNA level of RelA decreased from 1.00 ± 0.13 to 0.18 ± 0.00 (P<0.01); furthermore, the expression of RelA protein was weaken and the activited caspase-9 protein expression was increased gradually. Conclusions Micheliolide plays a significantly inhibited role in HeyA8, SKOV3 and A2780/DDP cells. The inhibited role of micheliolide inovarian cancer cells might through inhibiting nuclear factor-kappa B (NF-кB) signaling pathway, and inducing the expression of activited caspase-9 protein to promoting apoptosis of HeyA8 cell.

Objective To investigate the value of ADC histogram of tumor volume measurement in the diagnosis of prostate cancer,and to screen out the best diagnostic parameter value.Methods 31 cases of prostate cancer and 35 cases of benign prostatic hyperplasia confirmed by biopsy or surgical pathology were analyzed retrospectively.DWI examination was performed on all patients before treatment,and b value of 0 and 1500 s/mm2 was selected.The total tumor ADC histogram parameters were measured respectively,including the average value of ADC (ADCmean ),the median ADC (ADCmedian ),the tenth percentile ADC (ADC10th),the twenty-fifth percentile ADC (ADC25th), the fiftieth percentile ADC (ADC50th),the seventy-fifth percentile ADC (ADC75th),the ninetieth percentile ADC (ADC90th), skewness and kurtosis.The histogram parameters of the two groups of patinents and their diagnostic efficacy were analyzed and compared.Results ADCmean ,ADCmedian ,ADC10th,ADC25th,ADC50th,ADC75th and ADC90th in the prostate cancer group were statistically lower than those of benign prostatic hyperplasia group (P <0.01),and there was no significant difference in the skewness and kurtosis between the two groups of diseases (P > 0.05 ).ADCmean ,ADCmedian ,ADC10th,ADC25th,ADC50th,ADC75th and ADC90th diagnosing prostate cancer in the area under ROC curves (AUC)were more than 0.78.ADC10th had the best diagnostic efficacy and its AUC was 0.82, with the optimal cut-off value for 0.27 × 10 -3 mm2/s,with sensitivity and specificity for 78.4％ and 83.3％.Conclusion The ADC histogram of the total tumor volume measurement is of great value in the diagnosis of prostate cancer,among which ADC10th is the most effective parameter.It can accurately distinguish between prostate cancer and prostatic hyperplasia nodules.

Objective:To explore the dance movement therapy on nutritional status and muscle strength in older patients with sarcopenia.Methods:A total of 35 older sarcopenia patients from May 2019 to April 2020 in Nanfang Hospital, Southern Medical University were enrolled as the intervention group with dance movement therapy; another 35 older sarcopenia patients from May 2018 to April 2019 were selected as control group with routine exercise intervention. Nutritional indicators and muscle strength were compared between two groups on admission, at 1, 3, 6 months after intervention.Results:At 3, 6 months after intervention, hemoglobin and body mass index were (124.10 ± 16.59) g/L, (128.33 ± 14.50) g/L and (21.40 ± 1.87) kg/m 2, (21.40 ± 1.87) kg/m 2 in the intervention group, higher than in the control group (116.03 ± 12.23) g/L, (120.09 ± 11.34) g/L and (20.03 ± 1.93) kg/m 2, (19.97 ± 1.99) kg/m 2. The differences were statistically significant ( t values were 2.17-2.84, all P<0.05). At 6 months after intervention, serum albumin was (33.73 ± 5.23) g/L in the intervention group, significantly higher than that in the control group (30.88 ± 5.17) g/L, the difference was statistically significant ( t=2.16, P<0.05). At 3, 6 months after intervention, muscle strength were (21.63 ± 1.54) kg, (23.17 ± 1.72) kg and short physical performance battery scores were 7.83 ± 1.56, 7.47 ± 1.59 in the intervention group, significantly higher than in the control group (19.66 ± 2.50) kg, (20.91 ± 2.83) kg and 6.59 ± 1.64, 5.97 ± 1.49, the differences were statistically significant ( t values were 3.05-3.83, all P<0.05). At 6 months after intervention, appendicular skeletal muscle mass index (ASMI) was (6.03 ± 1.47) kg/m 2 in the intervention group, significantly higher than (5.13 ± 1.36) kg/m 2 in the control group, the difference was statistically significant ( t=2.52, P<0.05). Conclusions:Dance movement therapy can improve the nutritional status and muscle strength of older patients with sarcopenia.

Objective:To explore the values of albumin-bilirubin (ALBI) score, carcinoembryonic antigen (CEA) and combination of the two in the prognostic assessment of colorectal cancer patients with postoperative liver metastasis.Methods:The clinicopathological data of 98 colorectal cancer patients with postoperative liver metastasis who were admitted to Lianyungang Oriental Hospital and receiving adjuvant chemotherapy from January 2016 to March 2020 were retrospective analyzed. The data of serum protein, bilirubin, and CEA before chemotherapy were obtained, the relationship between serum protein and bilirubin was analyzed, and the ALBI score was calculated. The ALBI-CEA score was judged according to the ALBI score and the CEA level. ALBI score > -2.60 points was categorized as high ALBI group, and ALBI score ≤ -2.60 points was categorized as low ALBI group; CEA >5 ng/ml was categorized as high CEA group, and CEA ≤5 ng/ml was categorized as low CEA group; patients were categorized into 0, 1, and 2 points groups based on ALBI-CEA score. Overall survival (OS) and progression-free survival (PFS) of ALBI score, CEA and ALBI-CEA score subgroups were analyzed by Kaplan-Meier method; with the actual survival and progress status of the patients as the gold standard, receiver operating characteristic (ROC) curve was used to analyze the effect of 3 indicators to assess patients' OS and PFS, and area under the curve (AUC) was compared; Cox proportional hazards model was used to analyze the influencing factors of OS and PFS.Results:The median albumin and bilirubin levels of the 98 patients were 34.4 g/L (26.8-42.8 g/L) and 16.6 μmol/L (7.6-44.6 μmol/L), and the result of Pearson correlation analysis showed a negative correlation between the levels of albumin and bilirubin ( r = -0.282, P < 0.001). The 3-year OS and PFS rates in the high ALBI group were lower than those in the low ALBI group (OS rate: 9.2% vs. 33.3%, PFS rate: 7.7% vs. 18.2%), and the differences in OS and PFS between the two groups were statistically significant ( χ2 values were 27.64, 23.30, both P < 0.001). The 3-year OS and PFS rates in the high CEA group were lower than those in the low CEA group (OS rate: 7.1% vs. 42.9%, PFS rate: 7.1% vs. 21.4%), and the differences in OS and PFS between the two groups were statistically significant ( χ2 values were 23.71, 17.14, both P < 0.001). The 3-year OS rates in the ALBI-CEA score 0, 1 and 2 points groups were 77.8%, 20.9% and 2.2%, and the 3-year PFS rates were 44.4%, 9.3% and 6.5%, and there were statistical differences in OS and PFS among the three groups ( χ2 values were 102.36, 76.55, both P < 0.001). The ROC curve analysis showed that the AUC of ALBI score, CEA and ALBI-CEA score for assessing OS were 0.688 (95% CI 0.544-0.832), 0.754 (95% CI 0.618-0.890) and 0.828 (95% CI 0.723-0.933) (all P < 0.05), and the AUC for assessing PFS were 0.618 (95% CI 0.436-0.799), 0.646 (95% CI 0.464-0.829) and 0.682 (95% CI 0.494-0.870) (all P > 0.05). Multivariate Cox regression analysis showed that ALBI-CEA score was an independent influencing factor for OS (2 points vs. 0 point: HR = 17.254, 95% CI 8.385-35.504, P < 0.001) and PFS (2 points vs. 0 point: HR = 6.144, 95% CI 3.725-10.134, P < 0.001) of patients. Conclusions:The colorectal cancer patients with liver metastasis and high ALBI-CEA score are at high risk of death and disease progression and have a poor prognosis, and they are recommended to receive intensive treatment.

Objective To explore whether a single bout of exhaustive exercise can activate autophagy in mice skeletal muscles and to investigate the effect and possible mechanisms of exercise-induced autophagy on antioxidant defense function in vivo.Methods Thirty healthy 4-week-old male C57BL/6 mice were randomly divided into a control group(n=6) and an exercise group(n=24).The latter was subjected to a single bout of exhaustive treadmill exercise.Gastrocnemius muscles of 6 mice on both sides were then isolated right after,as well as 6h,12h and 24h after the running.The manganese super oxide dismutase(MnSOD),total superoxide dismutase(T-SOD) and Copper-Zinc superoxide dismutase (CuznSOD) activities were determined using the xanthine oxidase method and the content of the total anti-oxygen capability(T-AOC) using the colorimetric method.The expression of Beclin1,P62 and Bcl2 mRNAs,as well as their proteins were determined using the fluorescent quantitative PCR and Western blotting.Results The expression of Beclin1 mRNA in gastrocnemius muscles of the exercise group increased more significantly than that of the control group right after the exercise,as well as 6 hours later(P＜0.01).P62 mRNA expression also increased significantly(P＜0.01) 24h after the exercise.In contrast,the expression of P62 protein at 0h,12h and 24h after the exercise decreased more significantly(P＜0.05 or P＜0.01) than that of the control group.Similarly,the expression of Bcl2 protein at 0h,6h and 12h increased significantly(P＜0.05 or P＜0.01).The MnSOD and CuznSOD activity increased significantly right after the exercise(P＜0.05).The T-SOD activity decreased significantly(P＜ 0.01) 6h after the exercise.T-AOC content increased significantly 6h,12h and 24h after the exercise (P＜0.05 or P＜0.01).The P62 protein expression was negatively correlated with T-AOC content.Conclusion The single bout of exhaustive treadmill exercise can activate the expression of such autophagyrelated factors as Beclin1 mRNA and P62 protein and increase the autophagy to a certain extent to maintain antioxidant defensive function of the skeletal muscle.

Objective:To determine the median effective concentration (EC 50) of ropivacaine producing sensory-motor separation when used for femoral nerve block performed under ultrasound guidance. Methods:American Society of Anesthesiologists physical status Ⅰ-Ⅲ patients of both sexes, aged 18-64 yr, with body mass index of 18-30 kg/m 2, scheduled for elective arthroscopic knee surgery under general anesthesia, were enrolled in this study. Femoral nerve block was performed under ultrasound guidance.After successful location, ropivacaine 22 ml was injected with the initial concentration of 0.50%, and the ratio between the two consecutive concentrations was 1.1.The EC 50 of ropivacaine was determined by up-and-down sequential method.The positive response was considered as sensory block without motor block.The negative response was considered as sensory and motor block.When a negative response was found, the concentration was decreased in the next patient.When a positive response occurred, the concentration was increased in the next patient.Probit analysis was used to calculate the EC 50 and 95% confidence interval of ropivacaine producing sensory-motor separation when used for femoral nerve block which was performed under ultrasound guidance. Results:The EC 50 of ropivacaine producing sensory-motor separation was 0.186%, and the 95% confidence interval was 0.155%-0.205% when used for ultrasound-guided femoral nerve block. Conclusion:The EC 50 of ropivacaine producing sensory-motor separation is 0.186% when used for femoral nerve block performed under ultrasound guidance.