Objective To explore central mechanism of metformin(MET)in salt -sensitive hypertensive rats by assessing the effect of metformin on inflammation and oxidative stress in hypothalamic paraventricular nucleus (PVN),sympathetic nerve activity and blood pressure.Methods Eight -week -old male Dahl salt -sensitive rats were divided into 4 groups:the normal -salt diet control group[0.3% NaCl +intracerebroventricular(ICV)artificial cerebrospinal fluid(aCSF)],the normal -salt diet with MET group(0.3% NaCl +ICV MET 25μg/d],the high -salt diet control group (8% NaCl +ICV aCSF),the high -salt diet with MET group (8% NaCl +ICV MET 25μg/d). Mean arterial pressure(MAP)was determined every week by a tail -cuff occlusion.After 6 weeks,all rats were eutha-nized,and blood and brain tissues were collected.Then,the plasma norepinephrine(NE,an indicator of sympathetic activity)was detected by enzyme linked immune sorbent assay(ELISA).The expression levels of interleukin(IL)-1β,IL -10 and NOX -2[a subunit of NAD(P)H oxidase],superoxide dismutase(SOD)in the PVN were detected by immunofluorescence,immunohistochemistry and Western blot methods.Reactive oxygen species(ROS)was detec-ted by dihydroethidium(DHE)staining.Results The MAP level of high -salt diet with metformin group was attenu-ated compared with that of the high -salt diet control group[(129.55 ±6.52)mmHg vs.(154.47 ±6.57)mmHg, F =121.90,P <0.05].The change of plasma NE level of high -salt diet with metformin group was lower compared with that of the high -salt diet control group[(364.57 ±30.73)pg/mL vs.(547.68 ±25.08)pg/mL,F =179.24, P <0.05].The expression levels of IL -1β,IL -6,NOX -2 and ROS were markedly higher in high -salt diet with metformin than those of the high -salt diet control group(F =27.80,21.20,22.48,31.99,all P <0.05),which of IL -10 and SOD was lower(F =17.69,23.69,all P <0.05).Conclusion Metformin may attenuate blood pressure in salt -sensitive hypertensive rats,at least partly via decreasing inflammatory molecules and inhibiting oxidative stress in the PVN,subsequently inhibiting sympathoexcitation.

The purpose of this study is to evaluate the interaction effects of In-Chen-How (Artemisia capillaries Thunb.) on the pharmacokinetics of acetaminophen and on liver microsomal cytochrome P450 enzyme activity in rats. The rats were divided into control group (n=8) without In-Chen-How and the pretreated group (n=8) administered with In-Chen-How (approximately 1.0 mL·kg-1, according to weight) for 5 consecutive days. Rats in the control group received water simultaneously. Each rat was then given acetaminophen. The pharmacokinetic parameters of acetaminophen of the two groups were significantly different. In the In-Chen-How pretreated group, the maximum concentration of acetaminophen and the area under the plasma concentration-time curve were reduced about 58.4%, 56.7% and 55.4%. To further explain the results, liver microsomal suspensions were obtained from rats that were randomly divided into control and In-Chen-How pretreated group. The levels of CYP1A2 and CYP2E1 in hepatic microsomal protein from pretreated group were increased as compared to that from the control group. It indicated that In-Chen-How can stimulate the activity of CYP isozymes. The changes in the pharmacokinetics of acetaminophen resulting from the administration of In-Chen-How are related to an increase in metabolic activity of CYP1A2 and CYP2E1.

Two hundred women with singleton pregnancy at full term undergoing cesarean section with combined spinal-epidural anesthesia were randomly assigned into 2 groups (n=100 in each group). During anesthesia the polyvinyl chloride epidural catheter was used in control group and the wire-reinforced epidural catheter was used in study group.The results showed that the number of patients in whom blood or blood-tinted fluid was withdrawn from epidural catheter was significantly lower in study group than that in control group.The domestic wire-reinforced epidural catheter can reduce the incidence of blood vessel injury induced by insertion of epidural catheter.

Objective: To study the changes of andrographolides in the drying process of the extract of Andrographis paniculata Nees. Methods: HPLC method was applied to analyze andrographolide and 14 deoxy 11, 12 didehydroandrographolide in the process. Results: The content of andrographolide descended rapidly in the whole drying process, while the content of 14 deoxy 11, 12 didehydroandrographolide ascended at first 12 hours, declined in content was slowly to follow. Conclusion: Baking temperature is not the only main factor to stimulate the transformation of andrographolide and 14 deoxy 11, 12 didehydroandrographolide.

Objective To explore the interleukin (IL)-6,IL-10 and T cell subsets levels in bronchoalveolar lavage fluid(BALF) of children with refractory mycoplasma pneumonia.Methods A total of 53 children with refractory mycoplasma pneumonia were selected as the observation group,30 children with bronchial foreign body in our hospital were chosen as controls during the same period.ABC-double antibody sandwich ELISA method was used to detect IL-6,IL-10 levels and the CD3 +,CD4 + and CD8 + T levels were measured by means of flow cytometry in BALF.Results The IL-6 and IL-10 levels in BALF of children in the observation group were (63.25 ± 18.61) ng/ml,(31.83 ± 8.33) ng/ml respectively,and they were significantly higher than those of the controls[(30.51 ± 1.34) ng/ml,(11.01 ± 2.91) ng/ml] (P ＜ 0.05,respectively).The percentage of CD3 +,CD4 +,CD8 + T cells and the ratio of CD4 +/CD8 + T cells in BALF of the observation group were (48.47 ± 2.88)％,(21.16 ± 6.29)％,(23.04 ± 4.63)％,0.94 ± 0.33,respectively,and they were significantly lower than those of the controls [(64.24 ± 3.06) ％,(34.34 ± 7.59) ％,(26.71 ±5.29)％,1.56-±0.67] (P＜0.05,respectively).Conclusion The IL-6,IL-10 levels in BALF of children with refractory mycoplasma pneumonia significantly increased,suggesting that cell-mediated immunity play an important role in the pathogenesis of refractory mycoplasma pneumonia.

Objective:To explore the effect of digital rehabilitation system on the recovery of infants with cerebral palsy.Methods:Twenty-one children with cerebral palsy were treated with residual cerebral palsy in Putuo District,and 21 children with cerebral palsy were followed up.The patients were divided into two groups (n =21).The control group was treated by routine OT training by the parents,and the treatment group was treated with the digital rehabilitation system.Three months later,the efficacy was evaluated and compared.Results:After 3 months of treatment,the total effective rate (effective rate and effective rate) of the two groups was 90.5％ and 81％,respectively,and the treatment group was significantly higher than the control group (P ＜0.05).The PROM of the two groups was improved (P ＜0.01),and the PROM in the treatment group was significantly higher than that in the control group (P ＜0.05).FMFM was significantly higher than that before treatment (P ＜0.01),and FMFM was significantly higher in the treatment group than in the control group (P ＜0.01).Conclusion:Family (community) digital rehabilitation system can effectively improve the rehabilitation of children with cerebral palsy.

0.05).CONCLUSION:Ang-(1-7) inhibits the secretion of MCP-1 and ICAM-1 in cultured HUVEC in a concentration dependent matter induced by AngⅡ. The inhibitory effect of Ang-(1-7) may be through its specific receptor.

Objective: To explore the role and related mechanisms of angiotensin-(1-7)(Ang-(1-7)) on Toll-like receptor 4 (TLR4) mediated oxidized low-density lipoprotein(ox-LDL)-induced oxidative stress in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were cultured in vitro and divided into six groups: the control group (normal medium), the ox-LDL group(treated with 75 mg/L ox-LDL), the ox-LDL+ Ang-(1-7) group (1 μmol/L Ang-(1-7) pretreated for 30 minutes, then intervened with 75 mg/L ox-LDL), the ox-LDL+ Ang-(1-7)+ A-779 group(1 μmol/L A-779 (Mas receptor) pretreated for 30 minutes, 1 μmol/L Ang-(1-7) pretreated for 30 minutes, then intervened with 75 mg/L ox-LDL), the ox-LDL+ A-779 group (1 μmol/L A-779 pretreated for 30 minutes, then intervened with 75 mg/L ox-LDL), the ox-LDL+ HTA125 group (10 μg/L HTA125 (TLR4-blocking antibody) pretreated for 30 minutes, then intervened with 75 mg/L ox-LDL ). The corresponding index was detected after 24 hours after intervention. Apoptosis of cells were detected by Annexin V-FITC/PI double staining flow cytometry and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL). The generation of reactive oxygen species (ROS), products in oxidative stress, were detected by DCFH-DA staining. The mRNA and protein expression levels of NADPH oxidase 4(NOX4) and TLR4 were detected by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analysis respectively. Results: (1) The results of Annexin V-FITC/PI double staining flow cytometry showed that the proportion of apoptotic cells was higher in ox-LDL group than in control group ((21.18±1.40)% vs. (1.59±0.26)%, P<0.01), lower in ox-LDL+ Ang-(1-7) group((7.42±1.07)%) and ox-LDL+ HTA125 group((9.19±1.01)%) than in ox-LDL group (both P<0.01), higher in ox-LDL+ Ang-(1-7)+ A-779 group ((19.91±1.30)%) and ox-LDL+ A-779 group((20.47±0.95)%) than in ox-LDL+ Ang-(1-7) group (both P<0.01). (2) The TUNEL results showed that the proportion of apoptotic cells was higher in ox-LDL group than in control group((10.83±0.77)% vs. (2.83±0.82)%, P<0.01), lower in ox-LDL+ Ang-(1-7) group ((3.66±0.54)%)and ox-LDL+ HTA125 group((4.97±0.60)%) than in ox-LDL group(both P<0.01), higher in ox-LDL+ Ang-(1-7)+ A-779 group((10.69±0.62)%) and ox-LDL+ A-779 group((11.43±0.42)%) than in ox-LDL+ Ang-(1-7) group (both P<0.01). (3) ROS level was higher in ox-LDL group than in control group(0.093±0.014 vs. 0.053±0.011, P<0.01), lower in ox-LDL+ Ang-(1-7) group (0.063±0.011, P<0.01)and ox-LDL+ HTA125 group(0.070±0.010, P<0.05)than in ox-LDL group, higher in ox-LDL+ Ang-(1-7)+ A-779 group(0.088±0.003) and ox-LDL+ A-779 group(0.095±0.005) than in ox-LDL+ Ang-(1-7) group (both P<0.01). (4) The mRNA expression level of NOX4 was higher in ox-LDL group than in control group(11.74±0.65 vs. 1.00±0.00, P<0.01), lower in ox-LDL+ Ang-(1-7) group (2.85±0.75)and ox-LDL+ HTA125 group(5.57±0.52) than in ox-LDL group(both P<0.01), higher in ox-LDL+ Ang-(1-7)+ A-779 group(10.51±0.54) and ox-LDL+ A-779 group (11.04±1.01) than in ox-LDL+ Ang-(1-7) group (both P<0.01), higher in ox-LDL group than in control group(27.60±1.86 vs. 1.00±0.00, P<0.01), lower in ox-LDL+ Ang-(1-7) group (8.00±1.03)and ox-LDL+ HTA125 group(14.83±0.97)than in ox-LDL group(both P<0.01), higher in ox-LDL+ Ang-(1-7)+ A-779 group(24.81±2.19) and ox-LDL+ A-779 group (26.64±0.65)than in ox-LDL+ Ang-(1-7) group (both P<0.01). (5)The protein expression level of NOX4 was higher in ox-LDL group than in control group (0.61±0.09 vs. 0.23±0.02, P<0.01), lower in ox-LDL+ Ang-(1-7) group(0.27±0.03) and ox-LDL+ HTA125 group(0.22±0.02) than in ox-LDL group(both P<0.01), higher in ox-LDL+ Ang-(1-7)+ A-779 group (0.58±0.06)and ox-LDL+ A-779 group(0.61±0.03) than in ox-LDL+ Ang-(1-7) group (both P<0.01). The protein expression level of TLR4 was higher in ox-LDL group than in control group(0.18±0.02 vs. 0.08±0.01, P<0.01), lower in ox-LDL+ Ang-(1-7) group(0.07±0.01) and ox-LDL+ HTA125 group(0.09±0.01) than in ox-LDL group(both P<0.01), higher in ox-LDL+ Ang-(1-7)+ A-779 group(0.18±0.02) and ox-LDL+ A-779 group(0.20±0.02) than in ox-LDL+ Ang-(1-7) group (both P<0.01). Conclusion TLR4 mediated the ox-LDL induced injury in HUVECs, and Ang-(1-7) could attenuate ox-LDL induced injury in HUVECs by modulating the specific Mas receptors.

A15, a DNA aptamer with binding specificity for U87 glioma cells stably overexpressing the epidermal growth factor receptor variant III (U87-EGFRvIII), was generated by cell systematic evolution of ligands by exponential enrichment (cell-SELEX) using a random nucleotide library. Subsequently, we established a cell enzyme-linked assay (cell-ELA) to detect the affinity of A15 compared to an EGFR antibody. We used A15 as a detection probe and cultured U87-EGFRvIII cells as targets. Our data indicate that the equilibrium dissociation constants (K(d)) for A15 were below 100 nmol/L and had similar affinity compared to an EGFR antibody for U87-EGFRvIII. We demonstrated that the cell-ELA was a useful method to determine the equilibrium dissociation constants (K(d)) of aptamers generated by cell-SELEX.
Aptamers, Nucleotide ; metabolism ; Brain Neoplasms ; metabolism ; Cell Line, Tumor ; Glioma ; metabolism ; pathology ; Humans ; Mutation ; Protein Binding ; Receptor, Epidermal Growth Factor ; genetics ; metabolism ; SELEX Aptamer Technique ; methods

Objective:To establish the reference values of stimulation single fiber electromyography (SFEMG) in orbicularis oculi, and to explore its sensitivity in repetitive nerve stimulation (RNS) negative ocular myasthenia gravis (OMG) patients, and the relationship between jitter and various clinical parameters.Methods:Thirty-two healthy volunteers were included to establish the reference value of normal controls from January 2019 to December 2019. From December 2019 to January 2023, 36 OMG patients with negative RNS were collected. Quantitative MG score (QMGS) was performed, neostigmine test and antibody titers as well as thymus CT results were recorded. One side of the orbicularis oculi muscle was tested with a disposable concentric needle electrode in stimulation SFEMG, and the mean consecutive difference (MCD) value was calculated, which was compared with the average MCD value and upper limit of individual values in normal controls to evaluate whether the jitter was abnormal. Spearman correlation analysis of abnormal mean MCD values with QMGS and antibody titer was conducted.Results:Among the 32 healthy volunteers, there were 13 males and 19 females, the age was (46.8 ±18.7) years, and the MCD was (19.0 ±4.4) μs. The upper limit of the reference value was 27.7 μs for average MCD, and 37.4 μs for 10% individual values. Among 36 OMG patients negative at RNS tests, 20 were male and 16 were female, with a age of (37.2 ±17.0) years. The MCD was (29.9 ±14.7) μs, and Jitter was abnormal in 29 patients (81%). Among them, 20 (20/25) patients were antibody positive, 6 (6/26) patients had thymic hyperplasia, and 7 (7/26) patients had thymoma. The QMGS was 3(2, 4). There were 7 patients (19%) with normal jitter, whose QMGS was 3(2, 4). Among the patients with normal Jitter, 5 (5/5) patients were antibody positive, 2 (2/6) patients had thymic hyperplasia. There was no statistically significant difference in clinical indicators between the two groups of patients with abnormal or normal jitter. There was no significant correlation in antibody titer or QMGS with abnormal mean MCD value. Conclusions:The upper limit of the mean MCD value in the normal controls is 27.7 μs. The upper limit of a single value is 37.4 μs. Its sensitivity for OMG patients with RNS negative is 81%, and the abnormal mean MCD value does not show a significant correlation with various clinical indicators. Abnormal jitter indicates dysfunction of neuromuscular junction transmission, which is an important neuroelectrophysiological indicator for MG patients and is suitable for RNS negative patients. Orbicularis oculi muscle stimulation SFEMG provides a reliable and sensitive electrophysiological means for functional evaluation of neuromuscular junction.