Objective To evaluate the effects of different ratios of medicine dosage for isoflurane and propofol on postoperative cognitive function of rats with mild cognitive impairment (MCI).Methods Healthy male Wistar rats,aged 16-18 months,were used in the study.MCI model was established by occlusion of bilateral common carotid arteries.One hundred and fifty rats with MCI were divided into 5 groups (n=30 each) using a random number table:sham operation group (group S),isoflurane group (group I),propofol group (group P) and different ratios of medicine dosage for isoflurane and propofol groups (IP1,2 groups).The rats inhaled 1.9％ isoflurane for 3 h in group I.Propofol 40 mg · kg-1 · h-1 was infused intravenously for 3 h in group P.The rats inhaled 1.0％ isoflurane,and propofol 20 mg · kg 1 · h-1 was infused intravenously for 3 h in group IP1.The rats inhaled 1.4％ isoflurane,and propofol 10 mg· kg-1 · h-1 was infused intravenously for 3 h in group IP2.After disappearance of eyelash reflex,open reduction and internal fixation was performed after tibial fracture was induced.On 7 days after operation,contextual fear conditioning test and Y maze test were used to assess the cognitive function,and hippocampal tissues were obtained to count the viable neurons (using Nissl's staining) and CCAAT/enhancer-binding protein homologous protein (CHOP) and caspase-12 positive neurons (by immunofluorescence) in hippocampal CA1 region.Results Compared with group S,the rate of time spent in N arm,the rate of time spent freezing,and the number of viable neurons were significantly decreased,and the number of CHOP and caspase-12 positive neurons were significantly increased in IP2,I and P groups (P＜0.05),and no significant change was found in the parameters mentioned above in group IP1 (P＞0.05).Compared with group IP1,the rate of time spent in N arm,the rate of time spent freezing,and the number of viable neurons were significantly decreased,and the number of CHOP and caspase-12 positive neurons was significantly increased in IP2,I and P groups (P＜0.05).Compared with group IP2,the rate of time spent in N arm,the rate of time spent freezing,and the number of viable neurons were significantly decreased,and the nunber of CHOP and caspase-12 positive neurons was significantly increased in I and P groups (P＜0.05).Conclusion Combination of 1.0％ isoflurane and propofol 20 mg · kg-1 · h-1 does not aggravate the postoperative cognitive dysfunction of the rats with MCI.

OBJECTIVE:To establish a method for content determination of steroidal saponin in Allium sativum. METHODS:Pre-column derivatization of steroidal saponin was performed by using the derivatization agent of nitro-benzoic acid-chlorine. And HPLC was conducted to determine the content of steroidal saponin. The column was Shimadzu VP-ODS with mobile phase of aceto-nitrile-water mixed solution(80:20,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 254 nm,the column temper-ature was 25 ℃,and the injection volume of 20 μl. RESULTS:The linear rang of sarsasapogenin was 0-1.25 mg/ml(r=0.999 0);RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 93.1%-96.8%(RSD=1.56%,n=6). CON-CLUSIONS:The method is simple,stable with good separation,and can be use for the content determination of steroidal saponin in A. sativum.

0.05) or significantly enhanced(P0.05).In this case,the immunoadjuvant capability of CpG ODN was better than FQ68 genomes.Conclusion:CpG ODN or FQ68 genome enhances humoral and Th1 type of immune responses when packed into AIV oil emulsion vaccine,while the adjuvants do not show better adjuvanticity when separated from the oil emulsion vaccine.

OBJECTIVETo observe the effects of electroacupuncture (EA) at different acupoints on apoptosis-related serum and expression of microRNA (miRNA) in rats with myocardial ischemia, so as to explore its mechanism of action.

METHODSA total of 48 male Wistar rats were randomly divided into a normal group, a model group, a Neiguan group and a acupoint compatibility group, 12 rats in each group. Isoprenaline hydrochloride (ISO) with a daily dose of 2 mg/kg was subcutaneously injected for 14 days to establish the myocardial ischemia model in the model group, Neiguan group and acupoint compatibility group. Rats in the normal group were subcutaneously injected with an equal volume of normal saline. After modeling, rats in the Neiguan group were treated with EA at "Neiguan" (PC 6), while rats in the acupoint compatibility group were treated with EA at "Guanyuan" (CV 4), "Zusanli" (ST 36) and "Neiguan" (PC 6). Rats in the normal group and model group were treated with immobilization, once day for 21 days. The contents of creatine kinase-MB (CK-MB), vascular cell adhesion molecule-1 (VCAM-1) and endothelin-1 (ET-1) in serum were detected by enzyme-linked immunosorbent assay (ELISA); apoptosis index (AI) of myocardial cells was detected by TUNEL method; the expressions of miRNA-1, miRNA-133, miRNA-208 and miRNA-499 were detected by real-time PCR method.

RESULTSCompared with the normal group, the serum CK-MB, VCAM-1 and ET-1 were significantly increased in the model group, Neiguan group and acupoint compatibility group (all P < 0.01), and the apoptosis index was significantly increased (all P < 0.01). The CK-MB, VCAM-1 and ET-1 in the Neiguan group and acupoint compatibility group were significantly lower than those in the model group (all P < 0.01); the AI was reduced, which was more significant in the acupoint compatibility group (P < 0.05). Compared with the normal group, the expression of miRNA-133 was reduced (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly increased in the model group (all P < 0.01). Compared with the model group, the expression of miRNA-133 was increased (both P < 0.01) and that of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced (all P < 0.01) in the Neiguan group and acupoint compatibility group. Compared with the Neiguan group, the expression of miRNA-133 was increased (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced in the acupoint compatibility group (P < 0.01, P < 0.05).

CONCLUSIONEA at acupoints, especially acupoint compatibility group, could effectively prevent and treat myocardial ischemia, and the protective effect is possibly correlated to the double regulation on increasing the expression of miRNA-133 and inhibiting the expression of miRNA-1, miRNA-208, miRNA-499.

Acupuncture Points ; Animals ; Apoptosis ; Disease Models, Animal ; Electroacupuncture ; Endothelin-1 ; genetics ; metabolism ; Humans ; Male ; MicroRNAs ; genetics ; metabolism ; Myocardial Ischemia ; genetics ; metabolism ; physiopathology ; therapy ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Wistar

Objective To investigate the appropriate compatibility of appropriate compatibility of sevoflurane and propofol for patients with mild cognitive impairment (MCI) undergoing posterior lumbar interbody fusion in order to protect their cognitive function.Methods Eighty patients, 41 males, 39 females, aged 65-75 years, BMI 17-26 kg/m2, ASA physical status Ⅰ or Ⅱ, scheduled to undergo elective posterior lumbar interbody fusion, were to be scored according to Montreal cognitive assessment (MoCA), mini mental state examination (MMSE), dementia scale (CDR) and daily living ability scale (ADL) to identify patients with MCI before the surgery.They were randomly assigned to 4 groups (n=20 each) using a random number table: TCI propofol 2.0-2.5 μg/ml group (group P), TCI propofol 1.2 μg/ml+sevoflurane 0.6 MAC group (group PS1), TCI propofol 0.6 μg/ml+sevoflurane 0.9 MAC group (group PS2), 1.0-1.5 MAC sevoflurane group (group S).MoCA and MMSE were used to evaluate the cognitive function of patients 1 d before the operation (T0), after patients become wide-awake (T1), 3 d and 7 d after operation (T2 and T3).Apolipoprotein J (ApoJ) concentration related to cognitive function in blood samples, which were drawn at T0-T3 would be measured with ELISA method.Results Compared with T0, the scores of MMSE and MoCA in four groups decreased significantly (P＜0.05) at T1, the scores of MMSE and MoCA in group S decreased significantly (P＜0.05) at T2;compared with T1, the score of MMSE in the four groups increased significantly at T2, T3 (P＜0.05).The scores of MMSE at T1, T3 in group S decreased significantly compared with groups P, PS1 and PS2 (P＜0.05).The scores of MoCA at T2, T3 in group S decreased significantly compared with groups P, PS1 and PS2 (P＜0.05).Compared with T0, the concentration of plasma ApoJ in the four groups increased significantly at T1 (P＜0.05).Compared with T1, the concentration of plasma ApoJ in the four groups decreased significantly at T2 and T3 (P＜0.05).Compared with group PS1, the concentration of plasma ApoJ at T1, T3 increased significantly in groups S and group PS2 (P＜0.05).Conclusion TCI propofol 1.2 μg/ml combined with 0.6 MAC sevoflurane group is the appropriate compatibility of sevoflurane and propofol for patients with MCI undergoing posterior lumbar interbody fusion,because it has less negative influence on cognitive function and lower concentration of plasma ApoJ.

This study was aimed to evaluate effect of Qinghua Granules (QHG) on glycometabolism, pancreatic islet function and oxidative stress in type-2 diabetics with heat syndrome. A total of 60 cases of type-2 diabetics with heat syndrome (according to the Syndrome Element Syndrome Differentiation) were enrolled in the clinic of the Department of Endocrinology and Metabolism, Shuguang Hospital Affiliated to Shanghai University of Tradi-tional Chinese Medicine. The average age of enrolled cases was (57.9 ± 6.9) years. Enrolled cases were randomly divided into the treatment group and the control group. The original hypoglycemic plan was continued to use. In the treatment group, QHG was administrated. And in the control group, placebo was given. The administration dosage in both groups was one package per day. The treatment course was 12 weeks. The fasting and postpran-dial (120 min after standard meal) blood samples before and after medication were collected. The main evalua-tion indexes were fasting plasma glucose (FPG), postprandial plasma glucose (PPG) and hemoglobin A1c (HbA1c). The secondary evaluation indexes were homeostasis model assessment (HOMA2-%B, HOMA2-%S, HOMA2%-IR), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), maleic dialdehyde (MDA), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL) and low-density lipoprotein (LDL). The anal-ysis of variance was used in the comparison of efficacy between two groups . The results showed that HbA1c in the treatment group was obviously reduced, and HOMA2-%B was obviously increased. There was no significant changes in the control group ( P = 0 . 044 , P = 0 . 016 ) . In the treatment group , SOD increased obviously , MDA reduced obviously. There was no significant change in the control group. There was difference b etween two groups (P = 0.011, P = 0.049). There was no change on blood lipids or other evaluation indexes. It was conclud-ed that QHG is effective in the improvement of glycometabolism, islet β-cell functions and oxidative stress in type-2 diabetics with heat syndrome .

OBJECTIVETo explore the effects of principal-subordinate acupoints combination on improving myocardial ischemia, and the gene regulatory pathways for the protection of myocardial ischemia.

METHODSAccording to the random number table method, 70 SPF Wistar male rats were divided into a normal group, a model group, a LY294002 group, an insulin-like growth factors-1(IGF-1) group, a Neiguan group, an acupoint combination group and an acupoint combination + LY294002 group, 10 rats in each one. Rats in the normal group were injected with 0.9% NaCl solution, while rats in the remaining groups were treated with abdominal subcutaneous injection of isoroterenol hydrochloride to establish the rat model of myocardial ischemia. Rats in the LY294002 group and IGF-1 group were treated with injection of LY294002 solution and IGF-1 solution for 14 days. Rats in the Neiguan group were treated with electroacupuncture (EA) at "Neiguan" (PC 6) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination group were treated with EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination + LY294002 group were treated with LY294002 solution for 14 days, and EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) was given before model establishment, once a day for 21 days. EA pretreatment was given before model establishment in all acupuncture groups. The heart rate (HR) and ST segment voltage were detected before and after treatment; the myocardial pathological morphology was observed by HE staining; the expressions of P13K mRNA and Akt mRNA were tested.

RESULTSAfter modeling, HR and ST segment voltage in all intervention groups were higher than those in the normal group (all P < 0.01); after the intervention, the HR and the ST segment voltage in the acupoint combination group, IGF-1 group and IGF-1 group were improved (P < 0.01, P < 0.05), which was more significant in the acupoint combination group and Neiguan group (all P < 0.01). As for the myocardial pathological morphology, obvious myocardial ischemia was observed in the model group, and that in the LY294002 group was the most serious, and that in the acupoint combination+ LY294002 group was moderate. After intervention, the myocardial pathological damage in the IGF-1 group, Neiguan group and acupoint combination group was significant improved, which was more significant in the IGF-1 group and acupoint combination group. As for the expression of PI3K mRNA and Akt mRNA, compared with normal group, the expression of PI3K mRNA was increased in the remaining groups after modeling (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0. 01). The expression of Akt mRNA in the LY294002 group and acupoint combination + LY294002 group was reduced (P < 0. 01, P < 0.05), while that in the remaining groups was increased (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0.01).

CONCLUSIONThe principal-subordinate acupoints combination could improve heart rate and ST segment voltage in rats with chronic myocardial ischemia, reduce myocardial pathological damage, which is superior to single selection of "Neiguan" (PC 6). The PI3K/Akt signaling pathway may be involved in the regulation mechanism of principal-subordinate acupoints combination for the protection of chronic myocardial ischemia.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Chronic Disease ; therapy ; Electroacupuncture ; Electrocardiography ; Heart Rate ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Male ; Myocardial Ischemia ; enzymology ; pathology ; physiopathology ; therapy ; Myocardium ; pathology ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Rats ; Rats, Wistar

Objective To evaluate the effect of chidamide combined with matrine on proliferation and apoptosis of cutaneous T-cell lymphoma (CTCL) cell lines HH and Hut78,and to explore their apoptotic mechanisms.Methods Both HH and Hut78 cells were treated with 0.4 μmol/L chidamide and 0.6 g/L matrine alone or in combination for 24,48 and 72 hours,with those treated with dimethyl sulfoxide (DMSO) serving as control groups.MTS assay was performed to deteet cellular proliferation rates of HH and Hut78 cells at each time point.After 48-hour treatment,flow cytometry was conducted to detect cell apoptosis,and Western blot analysis to determine expression of apoptosis-related proteins in these cells.Statistical analysis was carried out by using repeated measures analysis of variance,one-way analysis of variance,and least significant difference (LSD)-t test for multiple comparisons.Results Compared with DMSO,chidamide and matrine alone or in combination could inhibit the proliferation of HH and Hut78cells to different extents (F =15.88,558.26,P ＜ 0.05,＜ 0.001,respectively).At 48 hours,the apoptosis rate in Hut78 cells was significantly higher in the matrine group (20.98％ ± 1.53％),chidamide group (22.44％ ± 7.74％) and combination group (44.53％ ± 1.85％) than in the control group (8.42％ ± 4.23％;LSD-t =4.76,5.31,13.69 respectively,all P ＜ 0.05),as well as in the combination group than in the matrine group and chidamide group (LSD-t =8.93,8.37 respectively,both P ＜ 0.01);no significant differences were observed in the apoptosis rate of HH cells between the matrine group (13.98％ ± 3.86％)or chidamide group (13.61％ ± 1.62％) and control group (11.44％ ± 1.43％,both P ＞ 0.05),while the combination group (20.94％ ± 0.64％) showed a significantly higher apoptosis rate compared with the control group,matrine group and chidamide group (LSD-t =7.37,5.40,5.69 respectively,all P ＜ 0.05).In the case of HH cells,the combination group showed significantly higher cleaved caspase-3 expression (all P ＜ 0.05),but significantly lower protein expression of E-cadherin,nuclear factor (NF)-κB,phosphorylated-Bad (p-Bad) and Bcl-2 compared with the other 3 groups (all P ＜ 0.05).In the case of Hut78 cells,the expression of E-cadherin,NF-κB,p-Bad and Bcl-2 was significantly lower in the matrine group,chidamide group and combination group than in the control group (all P ＜ 0.05),while cleaved caspase-3 expression was significantly higher in the chidamide group and combination group than in the control group (both P ＜0.05).No matter in HH cells or in Hut78 cells,there were no significant differences in Bad protein expression between the 4 groups (all P ＞ 0.05).Conclusion Chidamide in combination with matrine can inhibit proliferation and induce apoptosis of HH and Hut78 cells,likely by regulating the expression of apoptosis-related proteins E-cadherin,NF-κB,p-Bad,Bcl-2 and cleaved caspase-3.

AIM: To explore the expressive role of lipoprotein-associated phospholipase A_2, high sensitive C-reactive protein and matrix metalloproteinase-9 in vulnerable atherosclerotic plaques in a rabbit model. METHODS: Forty eight New Zealand white male rabbits were randomly divided into 4 groups (12 rabbits each): control group, stable plaque group, p53 group, and p53+drug group. Rabbits in control group were fed with a regular diet and underwent sham operation. Rabbits in stable plaque group, p53 group and p53+drug group underwent balloon induced arterial wall injury and then were fed on a diet with 1% cholesterol. The animals were all fed for 3 months, then the rabbits in p53 group and p53+drug group underwent Ad5-CMV p53 transfection at 10th week. Before killed, the animals in p53+drug group underwent pharmacological triggering with Russell's viper venom (RVV) and histamine to induce the rupture of the atherosclerotic plaques. At the 1st day and before sacrifice, the serum was collected for measuring Lp-PLA_2, hs-CRP, MMP-9, HDL, LDL and VLDL. The expressions of Lp-PLA_2, hs-CRP and MMP-9 in tissues were determined by the methods of hybridization and immunohistochemistry. RESULTS: At the end of 12th week, the serum and tissue levels of Lp-PLA_2 and MMP-9 in stable plaque group, p53 group and p53+drug group were significant different from those in control group and in each group at the first day (P<0.05). The serum levels of Lp-PLA_2 and hs-CRP in p53 group and p53+drug group were significantly higher than those in control group and stable group (P<0.05). The serum levels of Lp-PLA_2, hs-CRP and MMP-9 were all significantly different between p53 group and p53+drug group (P<0.05). At the end of 12th week, pathological results showed that 4 groups were normal artery, stable plaque, vulnerable plaque and rupture plaque, respectively. The fabric cap was thicker in plaque groups than that in normal group (P<0.05). The rupture and formation of thrombus were more significant in p53+drug group than those in p53 group. The serum level of Lp-PLA_2 had negative interrelated relationship with fabric cap in plaque groups (r=-0.710, P<0.01), and hs-CRP, MMP-9 had no interrelated relationships with fabric cap in plaque groups. CONCLUSION: Base on the successful establishment of the atherosclerotic plaque animal model, serum Lp-PLA_2 shows better interrelated relationships to plaques stability. Combination with hs-CRP and MMP-9, we can exactly evaluate the nature of plaques.

Objective:To determine the expression of glucose transporter 3 (GLUT3) in cutaneous squamous cell carcinoma (cSCC), and to evaluate its effect on the cSCC cell line A431.Methods:From June 2016 to December 2020, 22 paraffin-embedded tissue specimens were collected from patients with pathologically confirmed cSCC in the Department of Dermatology, Peking University Third Hospital, and 20 discarded normal skin tissues after dermatological surgeries served as controls. Immunohistochemical assay was performed to determine the GLUT3 expression in cSCC tissues and normal skin tissues. Cultured A431 cells were divided into two groups: GLUT3 overexpression group transfected with a lentiviral vector carrying the SLC2A3 gene, and negative control group transfected with an empty lentiviral vector. Real-time fluorescence-based quantitative PCR and Western blot analysis were conducted to determine the mRNA and protein expression of GLUT3 in A431 cells in different groups, the cell proliferation assay (MTS assay) was performed to estimate the cell proliferative activity, and the live-cell analysis system Incucyte S3 was used for real-time evaluation of the migratory and invasive abilities of A431 cells in different groups. The relative glucose consumption and lactic acid production in A431 cells at 48 hours were measured by using glucose and lactic acid assay kits, retrospectively. Two independent samples t-test was used for comparisons between two groups, and one-way analysis of variance was used for comparisons among multiple groups. Results:The GLUT3 expression was significantly higher in the cSCC tissues than in the normal skin tissues (immunohistochemical assay score: 9.39 ± 2.56 points vs. 2.30 ± 2.60 points; t = 8.91, P < 0.05). Compared with the negative control group, the mRNA and protein expression of GLUT3 markedly increased in the GLUT3 overexpression group. MTS assay showed significantly increased proliferative activity of A431 cells in the GLUT3 overexpression group compared with the negative control group after 24- and 96-hour treatment ( t = 2.49, 3.54, P = 0.048, 0.012, respectively); cell fusion rates in the scratched area were significantly higher in the GLUT3 overexpression group than in the negative control group in the cell migration assay at 6, 12 18, and 24 hours and cell invasion assay at 12, 18, and 24 hours (all P < 0.05). At 48 hours, the relative glucose consumption and lactic acid production in A431 cells were significantly higher in the GLUT3 overexpression group than in the negative control group ( t = 2.98, 2.20, P = 0.011, 0.038, respectively) . Conclusion:GLUT3 was highly expressed in the cSCC tissues, and may participate in the occurrence and development of cSCC by providing energy to cSCC cells via promoting glucose uptake in cells to enhance their proliferative, migratory and invasive abilities.