Effect of Yin Yang protein 1 transcriptionally regulating acetyl-CoA carboxylase 1 on the cell migration and lipogenesis in ovarian cancer / 肿瘤研究与临床

Objective:To investigate the expression of acetyl-CoA carboxylase 1 (ACC1) in ovarian cancer tissues and cells, and the related mechanisms of the effect of ACC1 on cell migration and lipogenesis in ovarian cancer.Methods:Samples including 1 case of normal ovarian tissue, 1 case of ovarian cancer primary lesion tissue and 1 case of ovarian cancer omentum metastatic tissue diagnosed by pathology examination of patients undergoing surgery resection who admitted to Linyi Cancer Hospital between January 2019 and December 2021 were collected. Immunohistochemistry was used to detect the protein levels of ACC1 and Yin Yang protein 1 (YY1) of all tissues. The PROMO database was used to predict the possible binding sites of YY1 and ACC1 promoter region. Through the assembled viral vector, the HEY cells of human ovarian cancer with ACC1 or YY1 expression [the untreated cells were treated as the negative control (NC)], or knocked down ACC1 or YY1 (the interference sequence sh1, sh2, sh3 was transferred to the target gene, and the negative control sequence shNC was transferred to the interference sequence). Double luciferase reporter gene assay was used to verify the binding sites of YY1 and ACC1 promoter and the activity of transcriptional regulation. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the mRNA and protein expression levels of ACC1 and YY1 in the treated HEY cells, respectively. Transwell assay was used to detect the migration ability of HEY cells. Oil red O staining and Nile red staining were used to detect the lipid droplets in HEY cells.Results:The immunohistochemical scores of ACC1 and YY1 were 0, 2, 8 scores and 0, 4, 6 scores, respectively in normal ovarian tissue, primary lesion of ovarian cancer, and omentum metastatic tissue. Transwell assay showed that the number of invasive HEY cells in ACC1 overexpression group was more than that in NC group [(87.7±7.4) vs. (52.2±4.2), t = 5.19, P = 0.003]. The number of invasive HEY cells in ACC1-sh1 group, and ACC1-sh2 group with the knockdown of ACC1 was less than that in shNC group [(21.2±1.5), (29.7±2.3) vs. (56.2±5.3); t value was 6.41, 3.77; P < 0.001, P < 0.005]. The number of lipid droplets in HEY cells in the ACC1 overexpression group was more than that in the control NC group [Oil red O staining: (301±25) vs. (215±21); Nile red staining: (287±15) vs. (207±10); all P < 0.05]; the number of lipid droplets in HEY cells in ACC1-sh1 and ACC1-sh2 group with the knockdown of ACC1 was less than that in ACC1-shNC group [Oil red O staining: (113±8), (119±12) vs. (195±18); Nile red staining: (82±8), (117±11) vs. (165±17); all P < 0.05]. The result of dual luciferase reporter assay showed that overexpression of YY1 promoted the luciferase activity of the wild type ACC1 promoter region report gene ( P = 0.003), while the luciferase activity of the report gene was inhibited compared with the wild type after the mutation of binding sites of YY1 in ACCI promoter region ( P = 0.008). Western blot results showed that the expression levels of YY1 and ACC1 protein in HEY cells with YY1 overexpression group were higher than those in NC group, which indicated a synergistic increasing trend of both YY1 and ACC1; the expression levels of YY1 and ACC1 protein in YY1-sh1 group, YY1-sh2 group and YY1-sh3 group with the knockdown of YY1 were lower than those in the control YY1-shNC group, which indicated a synergistic decreasing trend of both YY1 and ACC1. Conclusions:ACC1 and YY1 are highly expressed in ovarian cancer metastatic tissues and both show a positive correlation trend. The expression level of ACC1 in vitro has an impact on cell migration and lipogenesis in ovarian cancer via YY1 transcriptionally regulating ACC1.

Research progress of microRNAs in sepsis-associated acute kidney injury / 中华危重病急救医学

Sepsis is a life-threatening organ dysfunction syndrome caused by the host's maladjusted response to infection, and is one of the important causes of acute kidney injury (AKI). Sepsis-associated acute kidney injury (SA-AKI) has a high incidence, poor prognosis and high mortality. The pathogenesis of SA-AKI is very complex, and its pathogenesis has not been fully elucidated. Therefore, finding effective biomarkers for early diagnosis, treatment, disease development and prognosis of SA-AKI is an urgent clinical problem.

Low molecular weight heparin through TLR4/MyD88/NF-κB pathway protects PC12 cells against oxygen glucose deprivation induced inflammation / 中国医师杂志

Objective:To investigate the effect of low molecular weight heparin (LMWH) on the inflammatory response of PC12 cells induced by oxygen glucose deprivation (OGD) and its related mechanism.Methods:The PC12 cells were cultured in vitro were randomly divided into sham(control) group, OGD group, LMWH group and blocking agent group. The latter group was divided into six groups: Eritoran+ OGD group, LMWH+ Eritoran+ OGD group, ST2825+ OGD group, LMWH+ ST2825+ OGD group, pyrrolidinedithiocarbamate (PDTC)+ OGD group and LMWH+ PDTC+ OGD group. OGD cell model was established. Cell counting kit-8 (CCK-8) assay was used to detect cell activity. The expressions of toll-like receptor 4 (TLR4), MyD88 and nuclear factor κB (NF-κB) mRNA and protein were detected by real time polymerase chain reaction (qRT-PCR) and Western blot. The concentration of interleukin (IL)-1β, IL-6, tumor necrosis factor-α(TNF-α) and S100β were determined by enzyme linked immunosorbent assay (ELISA). Results:The cell activity of OGD group was significantly lower than that of control group on the first, second, third day ( P<0.05). Compared with OGD group, the activity of LMWH group was increased on the second, third day ( P<0.05), but lower than that of control group ( P<0.01). The mRNA expression of TLR4, MyD88 and NF-κB was significantly increased in OGD group compared with the control group ( F=144.9, F=710.5, 79.51, P<0.01). Compared with OGD group, the mRNA expression of TLR4, MyD88 and NF-κB were significantly decreased after treatment with LMWH ( P<0.01), and the specific inhibitor of TLR4, MyD88 and NF-κB enhanced the anti-inflammatory effect of LMWH. The protein expression of this pathway was consistent with that of the gene. The concentration of IL-1β, IL-6, TNF-α and S100β in OGD group was significantly higher than control group ( P<0.05). After treatment with LMWH, the concentrations of inflammatory factors and S100β were significantly decreased compared with OGD group ( P<0.01). When hinder TLR4, MyD88 and NF-κB respectively by Eritoran, ST2825 and PDTC, the concentrations of inflammatory factors and S100β were significantly decreased, but it was still higher than control group ( P<0.05). Conclusions:OGD can cause pathological damage of PC12 cells, including high expression level of S100β and aggravation of inflammatory reaction. LMWH can improve cell activity, down-regulate inflammatory reaction degree and protect the cells. Using inhibitors of TLR4/MyD88/NF-κB pathway to inhibit the corresponding target, the up-regulation of inflammatory factors by OGD can be inhibited in varying degrees. These suggested that LMWH may regulate inflammatory reaction of PC12 cells induced by OGD through TLR4/MyD88/NF-κB pathway.

Protective effect and mechanism of Ribociclib on sepsis induced-acute kidney injury / 中华危重病急救医学

Objective:To investigate the role of Ribociclib in sepsis induced-acute kidney injury (AKI) and its possible mechanisms.Methods:① Twenty adult male C57BL/6 mice were divided into sham operation group (Sham group; only open the abdomen without ligating or perforating the cecum, administered with sodium lactate buffer 12 hours before the sham operation), Ribociclib control group (administered with 150 mg/kg Ribociclib), cecal ligation and puncture (CLP) group (sepsis model induced by CLP; lactate buffer was given by intragastric administration 12 hours before CLP), and Ribociclib pretreatment group (administered with 150 mg/kg Ribociclib 12 hours before CLP) according to random number table, with 5 mice in each group. Kidneys were harvested 12 hours after the operation. Pathological changes in kidney were observed by hematoxylin-eosin (HE) staining. Tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) levels in mice kidney homogenate were measured by enzyme linked immunosorbent assay (ELISA). Western Blot was used to detect the expression of cell cycle-related protein phosphorylate retinoblastoma protein (p-Rb), apoptosis-related protein Bcl-2 and Bax. ② Mouse renal tubular epithelial (TCMK-1) cell line was used for in vitro experiment. The cells were divided into control group, Ribociclib group (treated with 5 μmol/L Ribociclib for 24 hours), lipopolysaccharide (LPS) group (treated with 200 mg/L LPS for 6 hours), Ribociclib+LPS group (replaced with the medium containing 5 μmol/L Ribociclib and 200 mg/L LPS for 6 hours after exposing with 5 μmol/L Ribociclib for 18 hours). Inflammatory cytokines in cell culture medium were detected by ELISA. The expression of p-Rb, Bcl-2 and Bax, autophagy-related proteins microtubule associated protein 1 light chain LC3b (LC3bⅡ, LC3bⅠ) and p62, phosphate protein kinase B (p-AKT), phosphorylated mammalian target of rapamycin (p-mTOR) were measured by Western Blot. Results:① Animal experiments showed that, compared with the Sham group, the kidney tissue of mice were significantly damaged, the levels of TNF-α and IL-6 were increased, the expressions of p-Rb and Bcl-2/Bax ratio were decreased in kidney tissue in CLP group; but there was no significant difference in indexes between Ribociclib control group and Sham group. Compared with the CLP group, kidney injury in mice pretreated with Ribociclib was significantly ameliorated, the pathological score was significantly decreased (1.48±0.16 vs. 2.68±0.16, P < 0.01), the levels of TNF-α and IL-6 in kidney homogenate were significantly decreased [TNF-α(ng/g): 340.55±34.96 vs. 745.08±58.86, IL-6 (mg/g): 17.33±1.01 vs. 114.20±20.49, both P < 0.01], the expression of p-Rb was furtherly decreased (p-Rb/β-tubulin: 0.14±0.01 vs. 0.73±0.06, P < 0.01), Bcl-2/Bax ratio was increased (0.89±0.06 vs. 0.62±0.10, P < 0.01). ② In vitro experiments showed that, compared with the control group, the releases of TNF-α and IL-6 were increased, the expression of p-Rb was decreased, the ratios of Bcl-2/Bax and LC3bⅡ/Ⅰ were decreased, the expressions of p62, p-AKT and p-mTOR were increased in LPS group; the expression of p-Rb was decreased after Ribociclib treatment in TCMK-1 cells. Compared with the LPS group, TNF-α and IL-6 were decreased [TNF-α (ng/L): 2.73±0.23 vs. 4.96±0.10, IL-6 (ng/L): 36.05±5.83 vs. 53.78±24.08, both P < 0.01], the expression of p-Rb was furtherly decreased (p-Rb/β-tubulin: 0.25±0.05 vs. 0.65±0.05, P < 0.01), the ratios of Bcl-2/Bax and LC3bⅡ/Ⅰ were increased (Bcl-2/Bax: 1.01±0.07 vs. 0.73±0.05, LC3bⅡ/Ⅰ: 2.08±0.31 vs. 1.04±0.01, both P < 0.05), the expressions of p62, p-AKT and p-mTOR were decreased (p62/β-tubulin: 0.59±0.01 vs. 1.09±0.08, p-AKT/β-tubulin: 0.61±0.03 vs. 1.20±0.06, p-mTOR/β-tubulin: 0.50±0.05 vs. 1.15±0.08, all P < 0.01) in the Ribociclib+LPS group. Conclusion:Ribociclib pretreatment ameliorated sepsis-induced AKI and AKT/mTOR pathway may be involved in the protective role of Ribociclib on kidney.

Neuroprotective effects and underlying mechanism of preconditioning with MK-801 in cerebral ischemia/reperfusion injury in rats induced by cardiac arrest-cardiopulmonary resuscitation / 中国医师杂志

Objective To explore effects of dizocilpine (MK-801) preconditioning on excitatory amino acids and inflammatory response in rats induced by cardiac arrest-cardiopulmonary resuscitation (CACPR).Methods 18 male Sprague Dawley (SD) rats were randomly divided into three groups:control group,CA group and CA + MK-801 group.To establish rat models of CA-CPR and keep samples of serum and specimens of brain tissues for following detection.The injury of neurons was observed by HE staining and expression of N-methyl-D-aspartic acid receptor (NMDAR) in brain tissues was detected by Western blot.The concentrations of interleukin 1 beta (IL-1 β) and tumor necrosis factor (TNF)-α in serum were detected by enzyme linked immunosorbent assay (ELISA).Results Neurons in CA group were disorganized,cells shrank,nuclei pyknosis,and cytoplasmic eosinophilia,accompanied by inflammatory cell infiltration.Preconditioning with MK-801 reduced the pathological damage of neuron and degree of macrophage infiltration.The relative expression of NMDAR protein in CA group were significantly higher than that in control group (907.9 ±24.9 vs 321.6 ± 18.4,P ＜0.001).Preconditioning with MK-801 significantly decreased the expression of NMDAR in CA + MK-801 group compared with that in CA group (512.4 ± 21.1 vs 907.9 ± 24.9).The CA group showed significantly increased concentrations of IL-1 β and TNF-α than that in control group (P ＜ 0.001),and this effect was abolished by preconditioning with MK-801.CA rats treated with MK-801 showed higher concentrations of IL-1 β and TNF-α than the control group.Conclusions Cardiac arrest causes pathological injury of neurons,up-regulates expression of NMDAR and aggravates inflammatory response.These results induce the apoptosis of nerve cells.Blocking glutamate receptor with MK-801 can inhibit expression of NMDAR,decrease level of cytokines,down-regulate inflammatory reaction degree therefore to protect the brain.

An experiment on the pathogenesis of early biotrauma in ventilator-induced lung injury / 中华危重病急救医学

Objective To investigate the pathogenesis of early biotrauma in ventilator-induced lung injury (VILI).Methods Twenty-four 8-week-old male specific-pathogen-free Sprague-Dawley (SD) rats weighing 250-300 g were randomly divided into sham group (S group), conventional mechanical ventilation group (L group) and high tidal volume (VT) mechanical ventilation group (H group) with 8 rats in each group. All rats received tracheostomy after anesthesia. Rats in S group received no mechanical ventilation but breathe room air spontaneously. All other parameters of the ventilator were the same in both mechanical ventilation groups, and the fraction of oxygen was set to 0.21, the rats in L group received 7 mL/kg VT, and those in H group received 28 mL/kg VT. Four hours after ventilation all rats were sacrificed and the lung tissues were harvested for wet/dry (W/D) ratio. Pathological injury score was evaluated by hematoxylin and eosin (HE) staining. Transferase-mediated deoxyuridine triphosphate-biotin nick end labeling stain (TUNEL) was performed to count the apoptosis cell in lung epithelial. Western Blot was performed to evaluate hemi-channel protein Pannexin-1 expression in lung homogenate. Bronchoalveolar lavage fluid (BALF) was collected, and the concentration of lactate dehydrogenase (LDH), isoprostane, adenosine triphosphate (ATP) and white cell count in BALF were measured. Yo-pro-1/propidium iodide (PI) double stain was performed to evaluate early apoptosis cell in BALF.Results There was no significant difference in lung injury between S group and L group. Compared with S group and L group, rats in H group showed significant lung injury, represented as alveolar rupture, inflammatory cell infiltration, interstitial edema and airway epithelial exfoliation, and the lung W/D ratio was increased significantly (5.1±0.2 vs. 4.4±0.2, 4.3±0.4, bothP< 0.01), pathological score was significantly increased [4.00 (4.00, 8.00) vs. 1.00 (0, 4.00), 2.00 (0, 4.75), bothP< 0.01], the white cell in BALF was significantly increased (×106/L: 2.97±0.46 vs.1.03±0.26, 0.79±0.19, bothP< 0.01), the level of LDH was significantly increased (U/L: 148.6±38.2 vs. 34.4±13.5, 78.6±13.9, bothP< 0.01), and the expression of Pannexin-1 in lung homogenate was significantly increased (Pannexin-1/GAPDH: 0.89±0.21 vs. 0.48±0.25, 0.61±0.17, bothP< 0.01), the ATP concentration in BALF was also significantly increased (nmol/L: 456.84±148.72 vs. 19.23±13.34, 113.26±57.90, bothP< 0.01). There was no significant difference in the apoptosis cell in lung tissue or the apoptosis cell rate, isoprostane level in BALF among the three groups [apoptosis cell in lung (cells/HP): 4.00 (3.00, 5.00) vs. 5.00 (4.00, 6.00), 4.00 (3.25, 6.00); apoptosis cell rate in BALF: (0.57±0.20)％ vs. (0.42±0.16)％, (0.58±0.19)％; isoprostane in BALF (μg/L): 3.85±0.46 vs. 3.83±0.60, 3.59±0.69, allP > 0.05].Conclusion The early pathogenesis of biotrauma in VILI is related to the release of inflammation mediator via membrane channel after activating by pressure stress, but not apoptosis and lipid peroxidation.

The clinical effect of superficial temporal artery-middle cerebral artery bypass combining with en-cephalo-duroarterio-synangiosis in the treatment of moyamoya disease / 中国医师杂志

Objective To explore clinical curative effects of superficial temporal artery-middle cer-ebral artery bypass combining with encephalo-duroarterio-synangiosis( STA-MCA+EDMS) in the treatment of moyamoya disease. Methods Thirteen cases were confirmed to be moyamoya disease by digital subtrac-tion angiography ( DSA) , and treated with STA-MCA+EDMS methods. Four cases were ischemic, and the others were hemorrhagic in this group. Results The follow-up was 6-12 months. The clinical symptoms were improved significantly, without cerebral ischemic attack in 4 ischemic cases. Seven hemorrhagic cases got blood from the external carotid artery by means of vascular bypass or temporal muscle, and established effective collateral circulation half a year after operation. Two hemorrhagic cases bleed again in July after operation. However, the family gave up treatment. Ten cases were good, one case was moderate, while two cases got bad. The total effective rate was 76. 9％. Conclusions STA-MCA+EDMS can improve cerebral hemodynamics of moyamoya disease, improve quality of life, and reduce incidence of stroke.

Protective effects of rapamycin induced autophagy on acute kidney injury induced by sepsis / 中华危重病急救医学

Objective To investigate the protective effect of autophagy inducer rapamycin on acute kidney injury (AKI) induced by sepsis. Methods Twenty-four Sprague-Dawley (SD) male rats were randomly divided into sham group, caecal ligation and puncture (CLP) model group, and rapamycin treatment group (Rap treatment group), with 8 rats in each group. The septic AKI model was reproduced by CLP in rats, and rats in sham group were given appendix isolation without ligation and puncture. The rats in Rap treatment group were given 1.6 mg rapamycin by intraperitoneal injection immediately after model reproduction, and the rats in CLP model group were injected with an equal amount of normal saline. The rats in all groups were sacrificed after collecting peripheral blood specimen at 24 hours after model reproduction, and the levels of blood urea nitrogen (BUN) and serum creatinine (SCr) were determined. The pathomorphology change in renal tissue was observed under light microscope after periodic acid Schiff (PAS) staining. Real-time polymerase chain reaction (real-time PCR, RT-PCR) was used to determine the mRNA expressions of renal tubular autophagy related molecules Atg-5 and Beclin-1. Western Blot was used to detect the expressions of renal tubular autophagy associated protein microtubule labeled protein 1 light chain 3-Ⅱ (LC3-Ⅱ) and Beclin-1 as well as apoptosis protein cytochrome C (Cyt C), Bax and Bcl-2. TdT-mediated dUTP nick-end labeling (TUNEL) assay was used to determine the renal tubular epithelial cell apoptosis. Results Rapamycin could alleviate pathomorphology changes in rats with septic AKI, and decrease the levels of BUN and SCr. Compared with sham group, the expressions of Atg-5, Beclin-1 and LC3-Ⅱ in CLP model group were significantly increased [Atg-5 mRNA (2-ΔΔCt): 2.34±0.04 vs. 1.00±0.03, Beclin-1 mRNA (2-ΔΔCt): 1.40±0.02 vs. 1.00±0.03, LC3-Ⅱ protein (gray value): 0.82±0.03 vs. 0.45±0.04, Beclin-1 protein (gray value): 0.59±0.06 vs. 0.29±0.03, all P < 0.01]. Rapamycin could further up-regulate the expressions of Atg-5, Beclin-1, and LC3 Ⅱ [Atg-5 mRNA (2-ΔΔCt): 3.28±0.19 vs. 2.34±0.04, Beclin-1 mRNA (2-ΔΔCt): 2.38±0.08 vs. 1.40±0.02, LC3-Ⅱ protein (gray value): 1.11±0.07 vs. 0.82±0.03, Beclin-1 protein (gray value): 0.85±0.05 vs. 0.59±0.06, all P < 0.01]. Compared with sham group, the apoptotic cells in CLP model group were increased significantly [(34.49±10.45)% vs. (2.78±1.40)%, P < 0.01], Cyt C and Bax protein expressions were significantly up-regulated (gray value: 0.87±0.02 vs. 0.46±0.03, 1.20±0.06 vs. 0.46±0.01, both P < 0.01), and Bcl-2 expression was significantly down-regulated (gray value: 0.64±0.02 vs. 1.33±0.09, P < 0.01). Rapamycin could effectively inhibit cell apoptosis [(15.44±5.50)% vs. (34.49±10.45)%, P < 0.01] and the protein expressions of Cyt C and Bax (gray value: 0.72±0.03 vs. 0.87±0.02, 0.84±0.03 vs. 1.20±0.06, both P < 0.01), and up-regulate the protein expression of Bcl-2 (gray value: 0.77±0.04 vs. 0.64±0.02, P < 0.01). Conclusion The protective effect of rapamycin on renal tissue of rat with AKI induced by sepsis was depended on cell apoptosis inhibition through inducing and promoting cell autophagy.

Effect of timing of initiation of renal replacement therapy on prognosis in septic patients with acute kidney injury：a Meta analysis / 中华危重病急救医学

ObjectiveTo systemically review the effects of timing of initiation of renal replacement therapy (RRT) on prognosis in septic patients with acute kidney injury (AKI).Methods A systematic search for randomized controlled trials (RCTs) and other clinical studies focused on comparing varied timing of initiation of RRT in septic AKI patients was performed in English or Chinese from PubMed, Web of Science, EMBASE, CNKI, Wanfang data, VIP from January 2000 to July 2014. Data screened were extracted with Cochrane systemically review method, and enrolled literature was collected for Meta analysis with RevMan 5.2 software. Total mortality, 28-day mortality, the total length of hospital stay and intensive care unit (ICU) stay in septic AKI patients with early or late initiation of RRT was analyzed. Funnel plots were drawn to detect publication bias.Results Five retrospective comparative studies with a total of 885 patients were enrolled. Random effect model in Meta analysis showed that there was no significant difference in total mortality between early RRT group and late RRT group [65.7% (226/344) vs. 68.7% (239/348), odds ratio (OR) = 0.71, 95% confidence interval (95%CI) = 0.38-1.31,P = 0.27]. The funnel plot demonstrated publication bias. Fixed effect model showed that there was significant difference in 28-day mortality between early RRT group and late RRT group [43.4% (66/152) vs. 55.3% (94/170),OR = 0.59, 95%CI = 0.36-0.94,P= 0.03]. The funnel plot demonstrated publication bias too. The data of total length of hospital stay and ICU stay could not be Meta-analyzed because of different data distribution, but no differences in total length of hospital stay and ICU stay between early and late RRT groups for septic AKI patients were found.ConclusionEarly RRT could reduce the 28-day mortality in patients with septic AKI compared with late RRT, but it did not lower the total mortality.

Efficacy ofα-Lipoic Acid Treatment on Sepsis-induced Acute Kidney Injury in Rats and Its Mechanisms / 中国医科大学学报

Objective To investigate the impact ofα?lipoic acid(ALA)treatment on sepsis?induced acute kidney injury in rats and explore the mechanisms. Methods A total of 32 male SD rats were randomized into 4 groups:normal control group(group A),ALA?treated control group (group B),sepsis group(group C)and sepsis with ALA treated group(group D). Group A and B underwent sham operation,while CLP operations were conducted in group C and D. Rats in both group B and group D were then administered with 200 mg/kg ALA by oral gavage immediately after the surgical procedure. Twenty?four hours after the surgical procedure blood samples were obtained for the evaluation of creatinine,BUN,TNF?α,IL?6 and IL?1β. Rat kidneys were rapidly removed for PAS stain. Western blot was employed to determine the expression of NF?κB. Results Pathologi?cal changes of kidney were induced by sepsis and the level of creatinine,BUN,TNF?α,IL?6 and IL?1βwere significantly increased by 178%,66%, 55%,114%and 110%(P<0.01). respectively;simultaneously the phosphorylation and nuclear expression of NF?κB p65 in kidney tissues were significantly increased by 144%and 102%(P<0.01). Sepsis?induced acute kidney injury also significantly reduced the expression of IκBαby 61%(P<0.01). These changes were significantly suppressed by early ALA treatment. Compared with C group,the level of creatinine,BUN,TNF?α,IL?6 and IL?1βwere significantly decreased by 48%,26%,25%,37%and 40%(P<0.05),respectively,and the relative expression of IκBαwas increased by 103%(P<0.05). Conclusion The present study demonstrated that ALA can suppress the activation of NF?κB,thus ameliorat?ing sepsis?related acute kidney injury.

The effects of fluid resuscitation on oxygenation index and prognosis in early stage of severe acute pancreatitis / 中华危重病急救医学

Objective To analyze the correlation between fluid equilibrium and oxygen index in patients at early stage (within 2 weeks) of severe acutepancreatitis (SAP),and to discuss the effects of fluid equilibrium after resuscitation on the prognosis.Methods A clinical study was conducted.Ninety-seven patients with SAP admitted into Shengjing Hospital of China Medical University directly or transferred into intensive care unit (ICU) in 24 hours after admission between March 201 1 to October 2013 were studied.Finally,65 patients were enrolled in statistical analysis,and those with termination of treatment prematurely were excluded.The patients received treatment protocol formulated by the same physician in ICU.Patients were divided into improved group and death group according to the outcome.The differences in fluid equilibrium on 1,2,3,7,14 days after admission of ICU between the two groups were compared.The correlation between fluid equilibrium and oxygen index was analyzed with curve fitting.Results Among 65 patients enrolled,53 of them were improved after intensive care and were transferred into ordinary wards.However,12 patients died in ICU.Patients in the improved group showed delayed positive fluid equilibrium,and some patients even showed negative fluid equilibrium.Patients in death group needed more fluid to achieve fluid equilibrium.There was a significant difference in the need of fluid to reach an equilibrium between improved group and death group [1 day:1 814.5 (905.2,2 152.8) vs.3 891.0 (2 524.2,5 714.5),Z=-3.303,P=0.001; 2 days:2 469.0 (1 456.0,3 696.0) vs.6 498.0(4 617.8,8 763.5),Z=-4.431,P＜0.001 ; 3 days:3 234.0 (1 098.0,4 295.5) vs.9 533.5 (6 748.8,10 689.0),Z=-4.684,P＜0.001 ; 7 days:3 234.0 (1 033.0,5 162.0) vs.13 986.5 (8 045.8,14 518.0),Z=-4.718,P＜0.001 ; 14 days:3 234.0 (978.5,4 924.0) vs.13 436.5 (8 045.8,14 518.0),Z=-4.769,P＜0.001].There was no correlation between fluid equilibrium and oxygen index in improved patients within 3 days of ICU admission (R 2=0.000,P=0.827),and it fit the logistic curve in a relatively low level after 3 days of ICU admission (R 2=0.036,P＜0.001).Conclusions Early fluid resuscitation could help maintain hemodynamics stability in SAP patients.Those SAP patients who showed a negative equilibrium in early stage showed a better prognosis,and the fluid equilibrium and oxygen index in improved patients fit the logistic curve after 3 days of ICU admission.

The effects of preconditioning and postconditioning with isoflurane on focal cerebral ischemi/reperfusion injury in rats / 中华危重病急救医学

Objective To investigate the effects of preconditioning and postconditioning with isoflurane on pro-inflammatory cytokines and lipid peroxidation in focal cerebral ischemic/reperfusion (I/R) injury in rats.Methods Thirty-two Sprague-Dawley (SD) rats were randomly divided into four groups:control group,model group,isoflurane preconditioning group and isoflurane postconditioning group,with 8 rats in each group.Rats in control group did not receive any challenge.In rats of model group right middle cerebral artery occlusion (MCAO) was conducted for 90 minutes.Rats in isoflurane preconditioning group received 2％ isoflurane exposure for 30 minutes 24 hours before MCAO for 90 minutes.Rats in isoflurane postconditioning group were given 60-minute 2％ isoflurane exposure after reperfusion of right MCAO.Twenty-four hours after the procedure,all rats were anesthetized with isoflurane,and blood sample taken from the heart was centrifuged,and the pro-inflammatory cytokines,including interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α),and lipid peroxidation products such as malonaldehyde (MDA) and superoxide dismutase (SOD) were determined.The mRNA and protein expression levels of matrix metalloproteinase (MMP-2,MMP-9),tight junction protein Calaudin-5 and Occludin were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot.Results Compared with control group,serum levels of IL-1 β (ng/L),TNF-α (ng/L) and MDA (μmol/L) were elevated and activity of SOD (U/L) decreased in rats of model group (IL-1β:76.81 ± 11.14 vs.52.43 ± 8.86,TNF-α:64.93 ± 10.81 vs.33.64 ± 7.94,MDA:8.63 ± 1.42 vs.4.14 ± 0.98,SOD:0.95 ± 0.21 vs.2.36 ± 0.80,all P＜0.05).After isoflurane preconditioning and postconditioning,compared with model group,the levels of IL-1 β,TNF-α and MDA were lowered,while activity of SOD was increased (IL-1 β:54.37 ± 9.06,56.82 ± 8.67 vs.76.81 ± 1 1.14,TNF-α:43.72 ± 6.16,39.49 ± 9.34 vs.64.93 ± 10.81,MDA:5.65 ± 0.83,5.82 ± 0.78 vs.8.63 ± 1.42,SOD:1.64 ± 0.47,1.71 ± 0.52 vs.0.95 ± 0.21,all P＜0.05).Focal cerebral I/R injury could lead to an increased expression of MMP accompanied with a decreased expression of tight junction protein.Compared with model group,after isoflurane preconditioning and postconditioning,it was found that there were decreased mRNA and protein expression of MMP-2 and MMP-9 (MMP-2 mRNA:1.25 ± 0.08,1.32 ± 0.12 vs.2.48 ± 0.26,MMP-2 protein:1.56 ± 0.09,1.50 ± 0.08 vs.2.12 ± 0.11 ; MMP-9 mRNA:1.26 ± 0.13,1.20 ± 0.12 vs.2.74 ± 0.28,MMP-9 protein:1.53 ± 0.04,1.51 ± 0.05 vs.2.23 ± 0.09,all P＜0.05) and increased levels of Calaudin-5 and Occludin (Claudin-5 mRNA:0.40 ± 0.08,0.38 ± 0.06 vs.0.28 ± 0.03,Claudin-5 protein:0.80 ± 0.06,0.81 ± 0.07 vs.0.39 ± 0.02; Occludin mRNA:0.54 ± 0.07,0.50 ± 0.08 vs.0.26 ± 0.06,Occludin protein:0.64 ± 0.06,0.69 ± 0.05 vs.0.49 ± 0.02,all P＜0.05).Conclusion Preconditioning and postconditioning with isoflurane can lower the levels of pro-inflammatory cytokines and the degree of lipid peroxidation,and lower the hydrolytic activity of MMP to the tight junction protein in cerebral tissue,thereby decrease the loss of tight junction protein and alleviate I/R injury.

Research progress on current pharmacokinetic evaluation of Chinese herbal medicines / 中国中药杂志

In order to prove safety and efficacy, herbal medicines must undergo the rigorous scientific researches such as pharmacokinetic and bioavailability, before they are put on the market in the foreign countries. Botanical Drug Products promulgated by the US FDA could guide industry sponsors to develop herbal drugs, which was also an important reference for investigating Chinese herbal medicines. This paper reviews and discusses novel approaches for how to assess systemic exposure and pharmacokinetic of Chinese herbal medicines, which were in line with FDA guidance. This mainly focus on identifying pharmacokinetic markers of botanical products, integral pharmacokinetic study of multiple components, Biopharmaceutics drug disposition classification system, and population pharmacokinetic-pharmacodynamic study in herb-drug interaction.

Transmembrane transport and metabolism of diammonium glycyrrhizinate across rat small intestine in Ussing Chamber / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the transmembrane transport and metabolism of diammonium glycyrrhizinate in intestines of rats.</p><p><b>METHOD</b>An Ussing Chamber model were used to investigate the transmembrane transport of diammonium glycyrrhizinate (GZ), the concentrations of diammonium glycyrrhizinate and its two metabolites were determined by HPLC.</p><p><b>RESULT</b>The permeability coefficients of GZ in difference intestinal mucous membranes were ranged from 0.3 x 10(-6) cm x s(-1) to 1.1 x 10(-6) cm x s(-1). The metabolism of GZ in enterocytes during its transport process was negligible. The concentration of diammonium glycyrrhizinate and pH had limit effects on the transport amount and the permeability coefficients of GZ.</p><p><b>CONCLUSION</b>GZ is a low permeability drug, but it can be absorbed at all segments of the small intestine in rats. Ileum is the major absorption region of GZ.</p>

Clinical study of the correlation of the serum precaldtonin level and the APACHE Ⅱ scoring in the patients of sepsis / 中国综合临床

Objective To evaluate the efficacy of serum PCT level in deciding the development and progno-sis of sepsis and its conrrelation with APACHE Ⅱ scoring.Methods 56 patients of sepsis accepted intensive care treatment and were all given APACHE Ⅱ scoring within the first 24 h after admission to ICU.The PCT level at dif-ferent time(1 d,3 d,5 d-7 d,10 d after admission)was detected.All these patients were divided into survival group and death group based on the 28-day fatality.Results The PCT level declined gradually with the treatment and it decreased obviously from the third day in comparison with the original level before admission [survival group/death group:(2.98±0.48)μg/L/(4.98±0.66)μg/L vs(4.04±0.50)μg/L/(6.02±0.50)μg/L](P<O.05).The PCT level in survival group declined quickly with the patients'condition improved and almost decreased to the normal level in the 10 day[0.48 ±0.18)μg/L],while the PCT in the death group was still in a higher level than normal even though it showed a tendency to decrease[(4.04±0.45)μg/L].The APACHE Ⅱ scores in death group was obviously higher than the survival group(death group/survival group:25.86±8.73/12.07±6.20,P<0·05).The coefficient of correlation between PCT and APACHE Ⅱ scoring was 0.656(P<0.05).Conclusion PCT,a single serology index,is characterized with conveniently and quick-which is strongly correlated with APACHE Ⅱscoring.

Study on chitosan and PHBHHx used as nerve regeneration conduit material / 生物医学工程学杂志

Both Chitosan and PHBHHx are natural, biodegradable biomedical materials. In this article, their ability to be made as nerve regeneration conduits are evaluated by studying their wettability, changes of the second structure of protein absorbed on their surface, growing status of fetal rat cerebral cortex nerve cells cultured on them, mechanical properties and ability to be processed later. The results indicate that both Chitosan and PHBHHx are promising nerve conduit materials.

Cloning of genes of Plasmodium berghei associated with benflumetol resistance / 军事医学科学院院刊

Objective:To isolate genes related to benflumetol resistance in the rodent malaria P.berghei K173 strain.Methods:The mRNA differential display technique was used to identify differentially expressed genes between drug-sensitive P.berghei K173 strain and benflumetol-resistant strain selected under benflumetol pressure.Northern blot was performed in order to determine the genes actually derived from maternal strains.Results:PCR reactions were prepared respectively with twenty-four pairs of primers in each of the strains.Thirty-three differentially displayed genes fragments were cloned and eight of them were sequenced. The eight sequences were novel ESTs and were submitted to GenBank. Homology searching revealed that clone CB52 was slightly homologous to P.falciparum cyclophilin gene. Hybridization confirmed that the gene fragment CB52 was upregulated in the benflumetol-resistant parasite strain, while downregulated in the chloroquine-resistant strain.Conclusions:CB52 fragment was involved in the benflumetol resistance in P.berghei and the genetic mechanisms of drug-resistance to benflumetol and chloroquine were different.

Therapeutic Efficacy of Naphthoquine Phosphate Combined with Artemisinine against Plasmodium knowlesi / 中国寄生虫学与寄生虫病杂志

Objective To study the antimalarial activity of naphthoquine phosphate combined with artemisinine against Plasmodium knowlesi in rhesus monkey.Methods Monkeys were randomly divided into 9 groups(3/group).The monkeys in groups A and B were treated i.g.once daily for 3 days with 6 or 10 mg/kg of naphthoquine phosphate respectively.Those in groups C and D were treated i.g.twice for the 1st day and once for the 2nd and 3rd day with 31.6 or 100 mg/kg of artemisinine respectively.In groups E, F and G, they were treated i.g.only once with the combination of naphthoquine phosphate 10 mg/kg and artemisinine 10, 20 or 25 mg/kg respectively.Groups H and I served as controls which were treated i.g.only once with 10 mg/kg of naphthoquine phosphate and 30 mg/kg of artemisinine respectively.Parasitemia was examined beginning 24 h after drug administration.The observation lasted 105 days when no more parasite was found.Results At 24 h after drug administration, the parasite reduction rate in all groups was higher than 90%.The parasite clearance time for groups E, F and G was(56.0?16.0),(53.3?4.6), and(56.0?8.0) h respectively, more rapid than that of Group H(69.3?4.6) h.There were 1, 3, 3, 2, 2, and 3 monkeys in groups A, B, D, E, F, and G respectively which were cured.No monkeys were cured in groups C, H and I.Conclusion The combination of naphthoquine phosphate and artemisinine is superior to the single component and the optimum proportion in the combination is 1:2.5 in treating P.knowlesi infection in monkeys.

Effects of transection of cervical sympathetic trunk on pregnant rats with L-NAME-induced hypertension and their fetuses / 中华麻醉学杂志

Bl on the 21st day of gestation. The difference was statistically significant ( P