Accuracy of serum pro-adrenomedullin concentration in predicting sepsis at different degrees of severity / 中华麻醉学杂志

Objective To evaluate the accuracy of serum pro-adrenomedullin (pro-ADM) concentration in predicting sepsis at different degrees of severity.Methods A total of 145 patients of both sexes,aged 18-64 yr,who were admitted to intensive care unit (ICU) of the First Hospital of Hebei Medical University from March 2015 to April 2017,with length of ICU stay＞24 h,were enrolled.The patients were divided into 3 groups according to the Sequential Organ Failure Assessment (SOFA) score within 24 h after admission to ICU:2＜SOFA score ≤ 6 mild sepsis group (n =50),6＜SOFA score ≤ 12 moderate sepsis group (n =50) and SOFA score＞ 12 severe sepsis group (n =45).Peripheral venous blood samples were collected immediately after admission to hospital for determination of serum pro-ADM concentrations by enzyme-linked immunosorbent assay.The receiver operating characteristic (ROC) curve of pro-ADM in predicting sepsis at different degrees of severity was plotted,and the area under the curve and 95％ confidence interval,cut-off value,sensitivity and specificity were calculated.Results The serum pro-ADM concentrations were significantly increased with the severity of sepsis,and the length of ICU stay was prolonged with the severity of sepsis (P＜0.05).In mild sepsis group,the area under the ROC curve was 0.770,95％ confidence interval 0.591-0.949,sensitivity 0.725,specificity 0.700 and cut-off value 6.45 nmol/L.In moderate sepsis group,the area under the ROC curve was 0.776,95％ confidence interval 0.645-0.907,sensitivity 0.813,specificity 0.760,cut-off value 8.30 nmol/L.In severe sepsis group,the area under the ROC curve was 0.83,95％ confidence interval 0.715-0.963,sensitivity 0.73,specificity 0.800,cut-off value 5.70 nmol/L.Conclusion Serum pro-ADM concentration can be used as a reliable index in predicting sepsis at different degrees of severity.

Effect of Clinically Equivalent Doses of Xuesaitong and Ginaton Injections on Cerebral Ischemia Reperfusion Injury of Rats / 医药导报

Objective To observe the curative effect of clinically equivalent doses of Xuesaitong and ginaton injections on cerebral ischemia reperfusion (I/R) injury of rats.Methods Male rats were randomly divided into five groups:normal control group,sham-operation group,model control group,Xuesaitong group and ginaton group.The cerebral ischemia rat model was established by middle cerebral artery occlusion (MCAO).Rats in the Xuesaitong group were given 20 mg·kg-1 of Xuesaitong injection,and rats in the ginaton group were intravenously injected with 7.5 mg· kg-1of ginaton immediately after I/R injury and once daily for 7 days.Rats in the sham-operation group and model control group were given the same volume of 0.9％ sodium chloride solution.The score of ethology,volume of cerebral infarction,mortality,superoxide dismutase (SOD),malondialdehyde (MDA),xanthine oxidase (XOD),nitrogen oxide (NO) and NO synthase (NOS) in seruu were examined.Results Compared with model control group,Xuesaitong and ginaton effectively reduced behavioral score 96 h (P ＜ 0.05),120 h (P＜0.01),144 h (P＜0.01) and 168 h (P＜0.01) after I/R injury,the volume of cerebral infarction 168 h after I/R injury and NO content (P ＜ 0.05).But they had no effects on NOS,SOD,MDA,and XOD contents.Conclusion Curatively injecting Xuesaitong and ginaton can effectively reduce cerebral I/R injury,but no significant difference in curative efficacy is observed between Xuesaitong and ginaton at clinically equivalent doses.

Value of serum CRISPLD2 levels for the diagnosis and prognosis evaluation of sepsis patients / 中华危重病急救医学

Objective To investigate the value of cysteine-rich secretory protein LCCL domain-containing 2 (CRISPLD2) in diagnosis and prognosis in patients with sepsis.Methods Clinical data of patients admitted to intensive care unit (ICU) of the First Hospital of Hebei Medical University from December 2014 to December 2016 were retrospectively analyzed. According to the severity of sepsis, the patients were divided into three groups: sepsis patients, severe sepsis patients and septic shock patients, and 100 healthy persons were enrolled as control group. Levels of serum CRISPLD2, procalcitonin (PCT) and C-reactive protein (CRP), acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score, sequential organ failure assessment (SOFA) score, and 28-day prognosis were recorded. Analysis of the correlation between CRISPLD2 and PCT, CRP, APACHEⅡscore, SOFA score was done. The receiver operating characteristic (ROC) curve was plotted for the CRISPLD2 value for the diagnosis and prognosis in patients with sepsis.Results A total of 115 patients with sepsis were enrolled in this study, including 52 sepsis, 48 severe sepsis, and 15 septic shock; 29 patients died after 28 days, 28-days mortality rate was 25.2%. There was no significant difference in CRISPLD2 between sepsis and healthy control group (mg/L: 204.1±74.5 vs. 211.3±12.0, P > 0.05); the level of CRISPLD2 in septic shock group was significantly lower than that in sepsis group and severe sepsis group (mg/L: 139.0±55.0 vs. 240.2±89.6, 233.0±8.9, bothP 216.0 mg/L, the sensitivity was 96.7%, and the specificity was 92.6%, which power lied between PCT and CRP. The AUC of CRISPLD2 for prognosis was significantly lower than that of PCT [0.617 (0.507-0.727) vs. 0.786 (0.668-0.903),P <0.01]; when the cut-off value of CRISPLD2 was 103.5 mg/L, the sensitivity was 100%, and the specificity was 25.6%. Conclusion CRISPLD2 is a potential biomarker in sepsis, but cannot predict the prognosis of patients with sepsis.

The clinical effect of airway pressure release ventilation for acute lung injury/acute respiratory distress syndrome / 中华危重病急救医学

Objective To evaluate the effect of airway pressure release ventilation (APRV) in patients with acute lung injury/acute respiratory distress syndrome (ALI/ARDS), to evaluate the extent of ventilator-induced lung injury (VILI), and to explore its possible mechanism. Methods A prospective study was conducted in the Department of Critical Care Medicine of the First Hospital of Hebei Medical University from December 2010 to February 2012. The patients with ALI/ARDS were enrolled. They were randomly divided into two groups. The patients in APRV group were given APRV pattern, while those in control group were given lung protection ventilation, synchronized intermittent mandatory ventilation with positive end-expiratory pressure (SIMV+PEEP). All patients were treated with AVEA ventilator. The parameters such as airway peak pressure (Ppeak), mean airway pressure (Pmean), pulse oxygen saturation (SpO2), mean arterial pressure (MAP), heart rate (HR), central venous pressure (CVP), arterial blood gas, urine output (UO), the usage of sedation and muscle relaxation drugs were recorded. AVEA ventilator turning point (Pflex) operation was used to describe the quasi-static pressure volume curve (P-V curve). High and low inflection point (UIP, LIP) and triangular Pflex volume (Vdelta) were automatically measured and calculated. The ventilation parameters were set, and the 24-hour P-V curve was recorded again in order to be compared with subsequent results. Venous blood was collected before treatment, 24 hours and 48 hours after ventilation to measure lung surfactant protein D (SP-D) and large molecular mucus in saliva (KL-6) by enzyme linked immunosorbent assay (ELISA), and the correlation between the above two parameters and prognosis on 28 days was analyzed by multinomial logistic regression. Results Twenty-six patients with ALI/ARDS were enrolled, and 22 of them completed the test with 10 in APRV group and 12 in control group. The basic parameters and P-V curves between two groups were similar before the test. After 24 hours and 48 hours, mechanical ventilation was given in both groups. The patients' oxygenation was improved significantly, though there were no significant changes in hemodynamic parameters. The Pmean (cmH2O, 1 cmH2O = 0.098 kPa) in APRV group was significantly higher than that in control group (24 hours: 24.20±4.59 vs. 17.50±3.48, P 0.05). The SP-D level (μg/L) in serum in APRV group showed a tendency of increase (increased from 19.70±7.34 to 27.61±10.21, P 0.05), the difference between the two groups was statistically significant (P 0.05). There was no significant difference in serum KL-6 between the two groups before and after ventilation. The SP-D and KL-6 levels in serum were unrelated with 28-day survival rate of the patients. The odds ratio (OR) of SP-D were 0.900 [95% confidence interval (95%CI) = 0.719-1.125], 1.054 (95%CI = 0.878-1.266), 1.143 (95%CI = 0.957-1.365), and the OR of KL-6 were 1.356 (95%CI = 0.668-2.754), 0.658 (95%CI = 0.161-2.685), 0.915 (95%CI = 0.350-2.394) before the test, 24 hours and 48 hours after ventilation (all P > 0.05). Conclusions APRV was similar to lung protective ventilation strategy in oxygenation and improvements in the lung mechanics parameters. APRV with a higher Pmean can recruit alveolar more effectively, and it had no impact on hemo-dynamics, but might exacerbate VILI.

Association of-429T/C and G1704 polymorphisms in the receptor for advanced glycation end products gene with proliferative diabetic retinopathy / 中华眼底病杂志

Objective To determine the association of-429T/C and G1704T polymorphisms in the receptor for advanced glycation end products gene with proliferative diabetic retinopathy (PDR).Methods Case-control study.From the Beijing Desheng Diabetic Eye Study cohort of 1467 patients with type 2 diabetes mellitus (T2DM),a total of 97 patients with PDR and 105 diabetic patients without retinopathy (DWR,duration of diabetes 15 years) were included for this study.Questionnaires were collected and general ophthalmologic examinations were performed.Biochemical analysis was conducted.DNA was extracted from peripheral venous blood.The-429T/C and G1704T single nucleotide polymorphisms were detected by the means of PCR-restrication fragment length polymorphisms.Results The frequency distribution of-429T/C in DWR group was 81.0％ in TT,16.1％ in TC,2.9％ in CC.The frequency distribution of-429T/C in PDR group was 77.3％ in TT,20.6％ in TC,2.1％ in CC.There was no significant statistical difference between the two groups (x2 =0.40,P＞0.05).Frequency of the-429T/C minor allele C in the DWR and PDR group were 11.0％ and 12.4％,respectively,with no significant statistical difference between the two groups (x2 =0.20,P＞0.05).The frequency distribution of G1704T in DWR group was 66.7％ in GG,29.5％ in GT,3.8％ in TT.The frequency distribution of G1704T in PDR group was 78.4％ in GG,21.6％ in GT.There was no significant statistical difference between the two groups (x2 =3.44,P＞0.05).Frequency of the G1704T minor allele T in the DWR and PDR group were 18.6 ％ and 10.8 ％,respectively,in which significant difference was found within the two groups (x2 =4.79,OR=1.88,95％CI:1.06-3.33,P＜0.05).Conclusions G1704T polymorphism is associated with PDR presence and 1704G allele may increase the risk of PDR.

Accuracy of vasopressin secretion in the late phase of septic shock for predicting patient outcomes / 中华麻醉学杂志

Objective To evaluate the accuracy of vasopressin (VP) secretion in the late phase of septic shock for predicting patient outcomes and further investigate its relationship with the prognosis of septic shock.Methods Fifty-five patients presented at late phase of septic shock,who were admitted to the intensive care unit of our hospital,were enrolled.Their VP secretion was measured.The method for measurement was as follows:3％ sodium chloride solution 600 ml was infused over 2 h,serum concentrations of VP and sodium were measured before and after infusion,the difference in VP before and after infusion (△VP) and in Na before and after infusion (△Na) was calculated,and △VP/△Na was used to reflect VP secretion.The patients were divided into either abnormal secretion of VP group (△ VP/△ Na ≤ 0.5 ng/mmol) or normal secretion of VP group (△VP/△Na＞0.5 ng/mmol) according to △VP/△Na ratio.Immediately before testing VP secretion,venous blood samples were collected for determination of serum lactic acid and C-reactive protein concentrations.The consumption of vasoactive drugs at the moment of enrollment and 28-day fatality rate were recorded.Results There were 30 cases in abnormal group (54％) and 25 cases in normal group (46％).Compared with normal group,the serum lactic acid,C-reactive protein concentrations and consumption of dopamine or norepinephrine were significantly increased,and the 28-day fatality rate was increased (67％ vs 40％) in abnormal group.ROC curve analysis showed that when △VP/△Na 0.5 ng/mmol was used as the criteria for determining prognosis,the sensitivity was 66.7％,specificity was 64.0％,and the area under the ROC curve was 0.828.Conclusion VP secretion in the late phase of septic shock may affect patient prognosis.

Effect of rhubarb on intestinal mucosal damage in septic rats: a comparison with ulinastatin / 中华麻醉学杂志

Objective To evaluate the effect of rhubarb on intestinal mucosal damage in septic rats by comparing with ulinastatin.Methods A total of 84 healthy Sprague-Dawley rats (half male,half female),aged 3 months,weighing 200-330 g,were randomly divided into 5 groups according to the random number table:control group (group C,n =6),sham operation group (group S,n =6),sepsis group (group Sep),rhubarb group (group R,n=24) and ulinastatin group (group U,n=24).Sepsis was induced by cecum ligation and puncture.In group R,rhubarb 1.2 g/100 g was dissolved in normal saline at room temperature,3 and 4 h later the filtrate about 2-3 ml was obtained and injected through a gastric tube into stomach once every 12 h,and 72 h later sepsis was induced.In group U,ulinastatin 50 000 U/kg (in 2 ml of normal saline) was injected once every 24 h,and 72 h later sepsis was induced.In Sep,R and U groups,at 6,12,24 and 48 h after ligation (T1 4),blood samples were collected from the orbital venous plexus for determination of plasma diamine oxidase (DAO) activity.Results The activity of plasma DAO was significantly higher at T1-4 in Sep,R and U groups than in C and S groups.The activity of plasma DAO was significantly lower at T3,4 in R and U groups than in Sep group.There was no statistical difference in the plasma DAO activity between R group and U group.Conclusion Rhubarb can reduce intestinal mucosal damage in septic rats,which is similar to that of ulinastatin.

Investigation of vasopressin response to increasing osmotic pressure in the late-phase of septic shock patients / 中华危重病急救医学

Objective To investigate the vasopressin (VP) response to increasing osmotic pressure in the late-phase of septic shock patients.Methods Thirty-seven septic shock patients hospitalized in intensive care unit (ICU) of the First Hospital of Hebei Medical Unive~ity from January 2012 to September 2013 were enrolled.All patients received 3％ hypertonic saline solution infusion.Serum concentrations of VP and sodium were measured before and after hypertonic saline solution infusion.Patients with ratio of difference in VP and sodium before and after infusion of 3％ hypertonic saline (△VP/△Na) ≤0.5 pg/mmol were defined as nonresponders,and who ＞0.5 pg/mmol defined as responders.The age,acute physiological and chronic health evaluation Ⅱ (APACHE Ⅱ) score,blood pressure,albumin level,vasoactive drug between the two groups were also analyzed.Results VP level in the nonresponsive group (n=20,54.05％) was markedly lowered before (ng/L:10.41 ± 1.70 vs.18.25 ± 5.90,t=5.29,P＜0.01) and after (ng/L:11.36 ± 1.90 vs.24.33 ± 5.46,t=9.33,P＜0.01) 3％ hypertonic saline solution infusion,compared with that in the responsive group (n =17,45.95％).All patients in the two groups were given dopamine (DA) or norepinephrine (NE) for maintaining blood pressure,and the dose in the nonresponsive group were higher than those in the responsive group [DA (μg· kg-1· min-1):14.91 ± 3.78 vs.8.64 ± 1.69,t =-5.02,P＜ 0.01 ; NE (μg· kg-1· min-1):1.03 ± 0.48 vs.0.38 ± 0.12,t=-3.12,P＜0.01].Three patients were given DA plus NE in the nonresponsive group while patients in the responsive group received only single drug therapy.The age,APACHE Ⅱ score,blood pressure,albumin level,sodium level before and after hypertonic saline solution infusion between the two groups were not statistically different.Conclusion VP secretion to osmotic challenge was impaired and decreased in the late-phase of septic shock,prompting dysfunction in VP synthesis.

Prognostic value of decreased vasopressin modulation in the late-phase of septic shock patients / 中华危重病急救医学

Objective To investigate the prognostic value of decreased vasopressin (VP)modulation in the late-phase of septic shock. Methods A prospective study was conducted. Fifty-five septic shock patients hospitalized in intensive care unit (ICU)of the First Hospital of Hebei Medical University from January 2012 to February 2014 were enrolled. All patients received 3% hypertonic saline solution infusion. Serum concentrations of sodium and VP were measured before and after hypertonic saline solution infusion. Patients with ratio of difference in sodium and VP before and after infusion of 3%hypertonic saline (△VP/△Na)≤0.5 pg/mmol were defined as non-responders,and who>0.5 pg/mmol were defined as responders. The levels of lactic acid,C-reactive protein (CRP),and vasoactive drug〔dopamine(DA)and norepinephrine(NE)〕usage between the two groups were compared. The 28-day mortality,live time in the dead,and ICU day in survivors were analyzed between the two groups. The receiver operating characteristic curve (ROC curve)was drawn to assess prognostic value of VP. Results There were 30 cases (54.5%) in non-responsive group,and 25 (45.5%)in responsive group. There were no significant differences in the age,acute physiology and chronic health evaluationⅡ (APACHEⅡ)score,central venous pressure (CVP),blood pressure, plasma albumin level,sodium level before and after hypertonic saline solution infusion between the two groups. The baseline level of VP in the non-responsive group was markedly lower than that of the responsive group (ng/L:10.66± 1.57 vs. 17.13 ±5.12,t=6.091,P<0.001). After hypertonic saline solution infusion,the VP level was also significantly decreased compared with that in the responsive group(ng/L:11.65±1.74 vs. 22.50±5.31,t=9.758,P<0.001). The non-responders showed higher lactic acid (mmol/L:3.04±0.55 vs. 2.28±0.38,t=-5.881,P<0.001) and CRP (mg/L:117.9±23.0 vs. 94.9±17.0,t=-4.143,P<0.001),and received larger dosage of vasoactive drugs〔DA(μg·kg-1·min-1):14.8±3.9 vs. 8.9±1.6,t=-5.725,P<0.001;NE(μg·kg-1·min-1):0.96±0.42 vs. 0.40± 0.09,t=-5.625,P<0.001〕for maintaining blood pressure compared with those in responders. The non-responsive group showed higher 28-day mortality(66.7%vs. 40.0%,χ2=3.911,P=0.048)and longer ICU day(days:9.9±2.3 vs. 6.7±1.7,t=-4.044,P<0.001),but the live time in the dead showed no difference between non-responsive group and responsive group(days:5.8±1.9 vs. 6.1±2.3,t=0.384,P=0.704). ROC curve showed that the area under ROC curve(AUC)forΔVP/ΔNa predicting the outcome was 0.828,and theΔVP/ΔNa threshold value of 0.5 pg/mmol had the sensitivity of 66.7%and specificity of 64.0%for prediction of the outcome(95%confidence interval:0.722-0.934). Conclusion Osmotic pressure-regulated VP secretion was impaired and decreased in the late-phase of septic shock, and made the sense in prognosis.

Safety and Efficiency of Anticoagulation Therapy during CVVH in the Patients with MODS and High-Risk Bleeding / 天津医药

Objective To investigate safety and efficiency of anti-coagulation therapy in patients with high-risk of bleeding and multiple organ dysfunction syndrome (MODS) during continuous veno-venous hemofiltration (CVVH). Meth-ods Forty patients with high-risk bleeding MODS during CVVH in our hospital were divided into heparin-free group (A group) and low-dose heparin group (B group). Blood coagulation function, platelets counts, blood urea nitrogen, serum creati-nine, PaO2/FIO2 and Apache Ⅱ scores in two groups were tracked before treatment and 24 h, 48 h after treatment. Filter lifespan, median ventilation time, ICU admission time and bleeding complications were observed. Results (1)There was significant difference in levels of blood urea nitrogen, serum creatinine, PaO2/FIO2 and ApacheⅡscores at 24 h, 48 h after treatment between in low-dose heparin group and those in heparin-free group (P＜0.05). (2)Levels of activated partial thromboplastin time(APTT), thrombin time (TT) were prolonged. Platelets count were significantly lower at 24 h after treat-ment than that before treatment in low-dose heparin group. Levels of APTT, TT and platelets count had no changes with pro-longed time of CVVH therapy.(3)Average ventilation time, ICU admission time were obviously shorter in low-dose heparin group than that in heparin-free group. Filter lifespan was significant longer in low-dose heparin group than that in heparin-free group, (P＜0.05).(4)Bleeding in skin and mucosa was observed in 1 case in low-dose heparin group without other se-vere bleeding complications. Conclusion The results of monocentric study show that low dose of heparin ensure smooth op-eration of CVVH in patients with MODS and high-risk bleeding. The clinical application is safe and efficient.

Inhibition of glutathione-S-transferase by total saponins of Panax notoginseng and its kinetics analysis in liver of mice / 中国中药杂志

<p><b>OBJECTIVE</b>To study the inhibition of glutathione-s-transferase by total saponins of Panax notoginseng and its kinetics analysis in liver of mice.</p><p><b>METHOD</b>Mouse liver cytochyma enzyme was obtained by different velocity centrifugation, the mouse liver glutathione-S-transferase of michaelis constant (Km), maximum velocity (Vmax) and the inhibition of glutathione-S-transferase by total saponins of P. notoginseng of 50% inhibiting concentration (IC50), inhibition constant (KI, KIS), the type of inhibition were calculation by Lineweaver-Burk and the low of semi-effet-probit.</p><p><b>RESULT</b>It was found that total saponins of P. notoginseng inhibited the glutathione-S-transferase activity with IC50 of 189.54 mg x L(-1). Kinetics analysis showed the glutathione-S-transferase of Km was 0.4563 mmol x L(-1) and Vmax was 476.19 U x mg(-1) with reduced glutathione (GSH) substrate, 0.1097 mmol x L(-1) (Km) and 400.00 U x mg(-1) (Vmax) with 1-chloro-2,4-dinitrobenzene (CDNB) substrate. Kinetics studies of total saponins of P. notoginseng on glutathione-S-transferase showed the inhibition were belong to mix-type with GSH and CDNB; the inhibition constant was 0.27 mg x L(-1) (KI), 0.68 mg x L(-1) (KIS) with GSH, and 0.21 mg x L(-1) (KI), 0.66 mg x L(-1) (KIS) with CDNB.</p><p><b>CONCLUSION</b>Total saponins of P. notoginseng strongly inhibited the glutathione-S-transferase activity.</p>

Effect of serum containing tengcha total flavonoid and dihydromyricetin on proliferation and apoptosis of HepG2 cells / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of serum containing Tengcha total flavonoid and dihydromyricetin on proliferation and apoptosis of HepG2 cells.</p><p><b>METHOD</b>Serum containing respectively Tengcha total flavonid, dihydromyricetins and CTX and control serum were prepared by serological pharmacology method. MTT assay was used to observe the proliferation inhibition rate of HepG2 cells after incubated with different kinds of serum. Inverted microscope was utilized to observe the morphological changes after HepG2 cells were treated with different serum. AnnexinV/7AAD double label method was used to detect earlier period apoptosis cells.</p><p><b>RESULT</b>Both serum containing 20% Tengcha total flavonid and serum containing 20% dihydromyricetin could restrain the HepG2 cells proliferation at different levels and the morpholological changes of apoptosis were observed. AnnexinV/7AAD double label method showed that the earlier period apoptosis cells rates were increased by serum containing 20% Tengcha total flavonoid, but serum containing 20% dihydromyricetin did not show influence on the earlier period apoptosis cells.</p><p><b>CONCLUSION</b>Tengcha total flavonoid can restrain the HepG2 cells proliferation and induce earlier period apoptosis cells.</p>

Expression of a protein elicitor pebC1 from Botrytis cinerea in Pichia pastoris / 生物工程学报

In order to express PebC1 in Pichia pastoris, the pebC1 sequence was amplified from genome Botrytis cinerea BC-4-2-2-1 by PCR and subcloned into the Pichua pastoris expression vector pPIC9K to generate pPIC9K-pebC1. The recombinant plasmid was linearized by Bgl II and transformed into Pichia pastoris GS115 by electroporation. Recombinant Pichia pastoris GS115/pPIC9K-pebC1 was screened by MD and G418-YPD plates and further confirmed by PCR. The protein expression was induced by methanol and analyzed by SDS-PAGE. SDS-PAGE analysis showed a special band about 39 kDa and western blotting indicated a good antigenicity of the expressed protein. Bioassay results showed that the recombinant protein PebC1 can induce resistance to gray mould disease of cucumber and Arabidopsi thaliana.

Extracellular expression of protein elicitor PeaT1 in Bacillus subtilis to enhance drought tolerance and growth in wheat / 生物工程学报

PeaT1, a protein elicitor from Alternaria tenuissima can promote plant growth and trigger systemic acquired resistance in plants. In order to expand the application of PeaT1, P43 promoter sequence and nprB signal peptide-encoding sequence were cloned from Bacillus subtilis 168 chromosomal DNA. The two sequences and peaT1 gene were spliced by overlapping extension. This product was cloned into the Escherichia coli-B. subtilis shuttle vector pHY300-PLK and the resultant recombinant expression vector (pHY43N- peaTI) plasmid was transformed into B. subtilis WB800. SDS-PAGE and Western blotting analysis showed that protein elicitor PeaT1 was expressed extracellularly in B. subtilis. This recombinant bacterial strain enhanced drought tolerance and promoted seedling growth in wheat.

Expression of peaT1 gene from Alternaria tenuissima in Pichia pastoris and its function / 生物工程学报

In this study, peaT1 gene was subcloned into the Pichia pastoris expression vector pPIC9K, which contained both the methanol-inducible promoter and the transcription terminator of the AOX1 gene, resulting the plasmid pPIC9K-peaT1. The recombinant plasmid was linearized by Sal I or Bgl II and transformed into P. pastoris GS115 by electroporation method. Recombinant strain was screened by Minimal Dextrose Medium and further confirmed by PCR. The gene was in frame integrated into the Pichia genome through homologous recombination, resulting the recombinant strain. Regulated by the alpha-Factor, promoter of AOX1 gene and termination signal of yeast genomic, the recombinant protein was expressed and secreted into the supernatant. The SDS-PAGE analysis indicated that the apparent molecular weight of target protein was about 35 kD. Bioassay results showed that the inhibition rate of the expressed protein against TMV was 30.37%. The supernatant was collected and then purified by anion exchange chromatography. This protein can promote seedling growth of wheat obviously.

Effect of Panax notoginseng on genes expression of CYP and GST in liver tissues of rats / 中国中药杂志

<p><b>OBJECTIVE</b>To examine the effects of Panax notoginseng on the expression of cytochrome P450 (CYP) genes and glutathione S-trans-ferase (GST) genes in liver tissues of male SD rats.</p><p><b>METHOD</b>Rats were administered P. notoginseng 2 or 4 g x kg(-1) bw/d by gavage daily for 14 days. The levels of gene expression of CYP1A1, CYP1A2, CYP2B1, CYP2E1, CYP3A1, CYP4A1, and GSTml, GST-pi were examined by quantitative real-time reverse-transcription polymerase chain reaction (quantitative real time-RT-PCR) assays.</p><p><b>RESULT</b>The expression of CYP1A1, CYP1A2, CYP2E1, CYP3A1, GSTml and GST-pi genes was not changed by 2 or 4 g x kg(-1) P. notoginseng treatment, But P. notoginseng significantly inhibited mRNA expressions of CYP2B1 (0.48-fold, P < 0.05, and 0.61-fold, P < 0.05, respectively) and CYP 4A1 (0.69-fold, and 0. 51-fold, respectively).</p><p><b>CONCLUSION</b>P. notoginseng had a special inhibitory selectivity on the expression of CYP2B1 and CYP4A1 genes in liver tissues of rats, which indicated it may be one of the mechanisms of actions of P. notoginseng. P. notoginseng had no effects on the expressions of CYP1A1, CYP1A2, CYP2E1 and CYP3A1 genes, which suggested when P. notoginseng co-administrated with those drugs metabolized by the above major metabolizing enzymes in liver, metabolic herb-drug interactions would not be happened.</p>

Effect of Panax notoginseng on genes expression of CYP and GST in lung tissues of rats / 中国中药杂志

<p><b>OBJECTIVE</b>To examine the effects of Panax notoginseng on the expression of cytochrome P450 (CYP) genes and glutathione S-transferase (GST) genes in lung tissues of male SD rats.</p><p><b>METHOD</b>Rats were administered P. notoginseng 2 or 4 g X kg(-1) bw/d by gavage daily for 15 days. The levels of gene expression of CYP1A1, CYP1A2, CYP1B1, CYP2B1, CYP2E1, CYP3A1, CYP4A1 and glutathione S-transferase ml (GST-ml) and glutathione S-transferase pi (GST-pi) were examined by quantitative real-time reverse-transcription polymerase chain reaction (quantitative real time-RT-PCR) assays.</p><p><b>RESULT</b>The expression of CYP2E1, CYP1A2 and GST-pi genes was not changed by P. notoginseng treatment, however, 2 g * kg-1 dose of P. notoginseng gave a 4.00-fold (P < 0.05) induction of CYP3A1 mRNA. P. notoginseng significantly increased mRNA expressions of GSTml (1.64-fold, P <0. 05 and 1.53-fold, P > 0.05) and CYP1A1 (3.44-fold, P > 0.05 and 6.05-fold, P < 0.05) in the 2 g x kg(-1) and 4 g x kg(-1) bw/d treatment groups, respectively, but P. notoginseng had a inhibitory tendency on mRNA expressions of CYP1B1 (0.81-fold, P > 0.05 and 0.38-fold, P > 0.05) and significantly inhibited the expressions of CYP2B1 (0.47-fold, P < 0.05 and 0.50-fold, P < 0.05) and CYP4A1 (0.54-fold, P < 0.05 and 0. 72-fold, P < 0.05) genes in the two groups.</p><p><b>CONCLUSION</b>A specific effect of P. notoginseng on the expression of different cytochrome P-450 genes or glutathione S-transferase genes in the lung tissues of rats was observed in this investigation. These findings would be very important and helpful for studying the mechanism of action of P. notoginseng and its reasonable use in clinic.</p>

Inhibition of glutathione-S-transferase by dihydromyricetin and its kinetics analysis in liver of mice / 中国药理学通报

Aim To study the inhibition of glutathione-S-transferase by dihydromyricetin and its kinetics analysis in liver of mice.Methods Mouse liver cytochyma enzyme was obtained by different velocity centrifugation,the mouse liver glutathione-S-transferase of michaelis constant(Km),maximum velocity(Vmax)and the inhibition of glutathione-S-transferase by dihydromyricetin of 50% inhibiting concentration(IC50),inhibition constant(Ki),the type of inhibition were calculated by Lineweaver-Burk and the low of semi-effect-probit.Results It was found that dihydromyricetin inhibited the glutathione-S-transferase activity with an IC50 of(121.14?13.66)?mol?L~-1.Kinetics analysis showed the Km was 0.1460 mmol?L~-1 and Vmax was 175.44 U?mg~-1 for reduced glutathione(GSH)substrate and 0.0937 mmol?L~-1(Km)and 212.77 U?mg~-1(Vmax)for 1-chloro-2,4,-dinitrobenzene(CDNB)substrate.Kinetics studies of dihydromyricetin on glutathione-S-transferase showed the inhibition was competitive with GSH and noncompetitive with CDNB,and the inhibition constant was 0.22 mmol?L~-1 with GSH and 0.54 mmol?L~1 with CDNB.Conclusion Dihydromyricetin can inhibit the glutathione-S-transferase activity in liver of mice.

EFFECT OF FERMENTATION CONDITIONS ON ANTIBIOTIC PRODUCTION OF XENORHABDUS NEMATOPHILUS / 微生物学通报

The optimum medium and fermentation conditions of the Xenorhabdus nematophilus from Steinernema carpocapsae BJ strain were studied. The relationship between antibiotic activity and pH, reducing sugar, total sugar, amino-nitrogen in process of fermentation was analyzed. The optimal medium contained tryptonl. 5 %, corn powderl o4, soybean flour 3 %, sucrose1 %KH2PO4 0.02 % ,MgSO4 0. 2% and activator 0. 1%, Stock cultured for 16h, inoculum size at 4%(V/V)and primary pH of medium ranged from 6.0 to 8.0, fermentation for 72h were of benefit to the yield of antibiotic. The pH, reducing sugar, total sugar and amino nitrogen in process of fermentation were related to the antibiotic activity. The yield of antibiotic increased by 56. 3 % comparison with nutrient broth.

EFFECT OF FERMENTATION CONDITIONS ON ANTIBIOTIC PRODUCTION OF XENORHABDUS NEMATOPHILUS / 微生物学通报

The optimum medium and fermentation conditions of the Xenorhabdus nematophilus from Steinernema carpocapsae BJ strain were studied. The relationship between antibiotic activity and pH, reducing sugar, total sugar, amino nitrogen in process of fermentation was analyzed. The optimal medium contained trypton1.5%, corn powder1%, soybean flour 3%, sucrose1%, KH 2PO 4 0.02%,MgSO 4 0.2% and activator 0.1%, Stock cultured for 16h, inoculum size at 4%(V/V)and primary pH of medium ranged from 6.0 to 8.0, fermentation for 72h were of benefit to the yield of antibiotic. The pH, reducing sugar,total sugar and amino nitrogen in process of fermentation were related to the antibiotic activity. The yield of antibiotic increased by 56.3% comparison with nutrient broth.