Objective To evaluate the efficacy and tolerance of antiretroviral therapy (ART)regimen containing lopinavir/ritonavir (LPV/r) as a second-line regimen.Methods Data of acquired immunodeficiency syndrome (AIDS) patients who have received the first-line therapy for over a year and changed to the second-line antiviral therapy regimen containing LPV/r for more than one year were collected retrospectively from the database of free antiviral therapy in Henan Province from January 1,2009 to December 31,2013.Based on the viral load inhibition status after the alteration of the regimen,the patients were assigned to virology failure with first-line therapy group,and successful viral inhibition but poor immunological reconstruction with first-line therapy group.The variation trend of CD4+ T lymphocyte counts of the two groups in 6,12,24 months after changed to the second-line regimen of LPV/r,the virology inhibition rates between 6 and 12 months,12 and 24 months,and occurrence of adverse events were analyzed.Quantitative data were analyzed by rank sum test,and qualitative data were analyzed by chi-square test.Results A total of 4 113 patients were divided into two groups,including the first-line therapy failure group (n=3 802) and poor immunological reconstruction group (n=311).At 6,12 and 24 months after the alteration of the regimen,the CD4+ T lymphocyte counts increased gradually (all P＜0.01).Between 6 and 12 months after the first-line therapy failure group changed to the secondline regimen,viral inhibition rate was 61.65％(1 408/2 284),while that 12 and 24 months was 68.91％(2 044/2 966).The incidences of adverse reaction of the two groups were 21.88％ (832/3 802) and 22.19％(69/311),respectively,which were not significantly different (x2 =0.015,P=0.901).Condusion The ART regimen containing LPV/r still has good viral inhibition effect and immunological reconstruction effect for AIDS patients who failed the initial therapy with poor immunological reconstruction.

Objective To investigate the feasibility of centerline measurement method in estimating aortic diameter at the proximal landing zone in Stanford B type aortic dissection. Methods CT angiography materials of 30 patients with type B aortic dissection were randomly selected from the hospital database (24 males with a median age of 49.5 years), which were retrospectively analyzed with multiplanar reformation (MPR) and centerline technique by two experts in vascular radiology. Difference between two measurement techniques was analyzed by using mixed linear model, and the agreement of measurements between two readers as well as between two techniques were evaluated by Bland-Altman plots. Results The diameters measured with MPR method by two experts were (29.73±2.99) mm and (29.86±2.95) mm respectively, while the diameters measured with centerline measurement method by two experts were (29.66 ±2.81) mm and (29.71 ±2.91) mm respectively. No statistically significant differences in the diameter value existed between the two measurement methods, although the results determined by centerline measurement method were more stable. Conclusion In determining aortic diameter at the proximal landing zone in Stanford B type aortic dissection, the centerline analysis provides a checking method for MPR measurement.

Objective To evaluate the virological and immune responses of Lopinavir/Ritonavir (LPV/r) based second-line regimen in elderly acquired immunodeficiency syndrome (AIDS) patients who failed first line regimens.Methods This was a retrospective cohort study.Elderly patients (≥50 years) who switched to LPV/r-based second-line antiretroviral therapy with human immuno-deficiency virus (HIV) RNA >1 000 copies/mL after more than 1 year of first-line treatment were recruited from Zhengzhou No.6 People Hospital from January 2010 to December 2011.The virological and immunological data during 60-month treatment were collected.Multivariate logistic regression was used to explore the risk factors associated with virological failure or immunological failure of 60-month second-line therapy.Results Totally 256 patients were enrolled with 109 male and 147 female.89.5% were plasma donator.The median age at the time of switching to LPV/r based second-line regimen was 61 years old.Twelve out of the 256 cases were detected for genotypic drug resistance and ten of them were resistant to drugs.No resistance to protease inhibitor (PI) was found.After switching to LPV/r based second-line regimen, HIV viral suppression (HIV RNA≤400 copies/mL) rates at 12, 24, 36, 48, 60 months were 69.5%, 78.4%, 79.0%, 79.7%, and 83.2%, respectively.The CD4+ T cell counts were (313±135) /mL at 12 months, (377±151) /mL at 24 months, (396±155) /mL at 36 months, (389±163) /mL at 48 months and (412±147) /mL at 60 months, which were all significantly higher than that at the initiation of therapy ([243±146] /mL,t=19.092,18.598,12.843,8.516 and 12.980, respectively;all P<0.05).After switching to LPV/r based second-line regimen for 60 months, 43 patients occurred virological failure and 48 patients occurred immunological failure.Multivariate logistic regression showed that poor adherence (OR=48.5, 95% CI: 15.9-98.4, P<0.01) and ART drug toxicity (OR=4.5, 95% CI: 2.6-11.3, P<0.01) were the main factors associated with virological failure at 60 months.Poor adherence (OR=15.1,95% CI: 6.89-33.3, P<0.01), CD4≤100 /mL at the time of switching therapy (OR=10.5,95% CI: 5.1-21.7, P<0.01), concomitant medications (OR=3.6,95% CI:1.6-4.1,P<0.01) were main factors associated with immunological failure at 60 months.Conclusions Elderly patients (≥50 years) who failed first line regimen should switch to LPV/r contained regimen as early as possible.Adherence education should be strengthened, drug toxicity as well as complications of treatment should be managed in time and concomitant medications should be reduced.

To synthesize aristolochic acid (AA)-2'-deoxyguanosine 5'-monophosphate (dGp) adducts in vitro and develop a novel method for the characterization of the adducts using multiple mass spectrometric techniques. AA was incubated with dGp in vitro using either enzymatic activation (by xanthine oxidase) or chemical activation (by zinc) to synthesize AA-dGp adducts, and the reaction conditions were optimized. Crude extracts were analyzed by techniques of liquid chromatography-electrospray ionization/tandem mass spectrometry (LC-MS/MS) and high accuracy mass data and isotope pattern of super high resolution Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICRMS). The quasi-molecular ion peaks of the AA-dGp adducts were obtained in the negative ion mode. Analysis by electrospray ionization/tandem mass spectrometry (ESI-MS/MS) provided useful structural information about AA-dGp adducts. AA can bind covalently to the exocyclic amino group of deoxyguanosine to form AA-dGp adducts. MS analysis is a powerful tool to detect and identify AA-dGp adducts simply, rapidly and accurately.
Aristolochic Acids ; chemical synthesis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; DNA ; chemistry ; metabolism ; DNA Adducts ; chemical synthesis ; Deoxyguanosine ; chemistry ; Tandem Mass Spectrometry ; methods

Objective To evaluate the feasibility and safety of hypothermic perfusion via renal artery balloon catheter before laparoscopic partial nephrectomy.Methods A total of 35 patients,who were arranged to receive laparoscopic partial nephrectomy during the period from March 2013 to December 2016 at the General Hospital of PLA,China,were enrolled in this study.The tumor was located in the left kidney in 22 patients and in the right kidney in 13 patients.The long diameter of the tumors was 2.3-7.0 cm,with a mean of (4.1±1.2) cm.Before laparoscopic partial nephrectomy,the implantation of renal artery balloon catheter was performed in all patients.The intraoperative renal cold-ischemia time,the time spent for operation and the amount of intraoperative blood loss were recorded.The creatinine clearance rate (Ccr) and estimated glomerular filtration rate (eGFR) were determined before operation as well as at one,3 and 7 days after operation.The intraoperative and postoperative complications were analyzed.Results Successful implantation of renal artery balloon catheter was accomplished in all 35 patients,with the technical success rate being 100％.Slipping of balloon catheter before laparoscopic partial nephrectomy occurred in one patient;in 2 patients incomplete occlusion of renal artery was found during operation and artery forceps had to be used to occlude the renal artery.In the other 32 patients,laparoscopic partial nephrectomy under the condition of hypothermic perfusion via renal artery balloon catheter was successfully carried out.No severe complications occurred during and after the operation.The average cold-ischemia time of the diseased kidney was 45 (20-125) min,the mean time for interventional procedure was 28 (20-40) min,the average surgery time was 147(95-235) min,the average amount of intraoperative blood loss was 180(50-1000) ml.The Ccr values determined before and at one,3,7 days after nephrectomy were (96.5±15.1),(75.2±10.5),(54.3±13.8) and (91.8±14.1) ml/min respectively.The eGFR values determined before and at one,3,7 days after nephrectomy were (99.5±15.3),(70.3±12.5),(65.5±11.7) and (96.8±12.3) ml·min-1·m-2 respectively.Statistical analysis indicated that in both groups the preoperative and 7-day postoperative Ccr values and eGFR values were significantly different from their one-day and 3-day postoperative values (P＜0.01),but no statistically significant differences existed between preoperative values and 7-day postoperative ones (P＞0.05).Conclusion Hypothermic perfusion via renal artery balloon catheter before laparoscopic partial nephrectomy is clinically safe and feasible,it can prolong the safe time of renal ischemia and protect renal function.

Objective To investigate the effect of silencing E6-associated protein(E6AP)on the level of p53 protein in human papilloma virus(HPV)negative cervical cancer cells(C33A cells).Methods The siRNA sequence silencing E6AP(siE6AP)and silencing control disordered siRNA sequence(siControl)were transfected into C33A cells with the mediation of LipofectamineTM2000 transfection reagent respectively.The silencing effect of siRNA on E6AP and the expression of p53and cleaved-caspase-3 proteins were detected by Western blot.Results The levels of E6AP protein in C33A cells of siE6AP group were significantly lower(t =-4.597,P<0.05),while the levels of p53 and cleaved-caspase-3 proteins were significantly higher than those of siControl group(t = 4.533 and 7.099 respectively,each P<0.05).Conclusion Silencing of E6AP significantly increased the expression of p53 protein in C33A cells,indicating that silencing of E6AP may restore the activity and function of p53 protein in C33A cells.

Objective To study the effect of glutamate on metabolism, shifts in glycolysis and lactate release in rat astrocytes.
Methods After 10 days, secondary cultured astrocytes were treated with 1 mmol/L glutamate for 1 h, and the oxygen consumption rates (OCR) and extra cellular acidification rate (ECAR) was analyzed using a Seahorse XF 24 Extracellular Flux Analyzer. Cell viability was then evaluated by MTT assay. Moreover, changes in extracellular lactate concentration induced by glutamate were tested with a lactate detection kit.
Results Compared with the control group, treatment with 1 mmol/L glutamate decreased the astrocytes’ maximal respiration and spare respiratory capacity but increased their glycolytic capacity and glycolytic reserve. Further analysis found that 1-h treatment with different concentrations of glutamate (0.1-1 mmol/L) increased lactate release from astrocytes, however the cell viability was not affected by the glutamate treatment.
Conclusion The current study provided direct evidence that exogenous glutamate treatment impaired the mitochondrial respiration capacity of astrocytes and enhanced aerobic glycolysis, which could be involved in glutamate injury or protection mechanisms in response to neurological disorders.

Objective To investigate the clinical value of vascular ultrasound in evaluating patients with unilateral symptomatic internal carotid artery occlusion(ICAO)before and after revascularization.Methods A retrospective analy-sis was performed for 45 patients who underwent revascularization due to unilateral symptomatic ICAO in The Second Hos-pital of Lanzhou University from January 2021 to July 2024,and according to the surgical procedure for revascularization,they were divided into carotid artery stenting(CAS)group with 32 patients and carotid endarterectomy(CEA)group with 13 patients.The two groups were compared in terms of general data,revascularization rate,change in the inner diameter of diseased vessels after surgery,opening of collateral circulation,and changes in hemodynamic parameters of the affected middle cerebral artery(MCA)after surgery.Results The revascularization rate was 84.4%for all 45 patients,and there was no significant difference in revascularization rate between the CAS group and the CEA group(87.5%vs 76.9%,χ2=0.188,P=0.665).Both groups had a significant change in the inner diameter of the distal internal carotid artery after treatment(the CAS group:t=-5.217,P<0.001;the CEA group:t=-5.056,P<0.001).There were no significant differ-ences between the two groups in the number and type of collateral opening before surgery(all P>0.05).For both groups,systolic blood flow velocity and pulsatility index of the affected MCA returned to normal after surgery(P<0.001).Conclusion Vascular ultrasound can be used to evaluate the changes in the structure,hemodynamic parameters,and collateral circulation of diseased vessels after revascularization in patients with unilateral symptomatic ICAO,and it is a simple and reliable evaluation method before and after revascularization.

The rhizome of Alpinia officinarum is a widely used Chinese herbal medicine. The essential oil in A. officinarum rhizome is mainly composed of 1, 8-cineole and other monoterpenes, as the major bioactive ingredients. In plants, monoterpenes are synthesized through the methylerythritol phosphate (MEP) pathway in the plastids, and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) is an enzyme catalyzing a committed step of the MEP pathway. In the present study, the full-length cDNA encoding DXR was cloned from the rhizome of A. officinarum, using homology-based RT-PCR and rapid amplification of cDNA ends (RACE) techniques. The new cDNA was designated as AoDXR and submitted to GenBank to be assigned with an accession number HQ874658. The full-length cDNA of AoDXR was 1 670 bp containing a 1 419 bp open reading frame encoding a polypeptide of 472 amino acids with a calculated molecular mass of 51.48 kDa and an isoelectric point of 6.15. Bioinformatic analyses revealed that AoDXR showed extensive homology with DXRs from other plant species and contained a conserved plastids transit peptide, a Pro-rich region and two highly conserved NADPH-binding motifs in its N-terminal region characterized by all plant DXRs. The phylogenetic analysis revealed that AoDXR belonged to angiosperm DXRs. The structural modeling of AoDXR showed that AoDXR had the typical V-shaped structure of DXR proteins. The tissue expression pattern analysis indicated that AoDXR expressed strongly in leaves, weak in rhizomes of A. officinarum. Exogenous methyl jasmonate (MeJA) could enhance the expression of AoDXR and the production of 1, 8-cineole in A. officinarum rhizomes. The cloning and characterization of AoDXR will be helpful to reveal the molecular regulation mechanism of monoterpene biosynthesis in A. officinarum and provides a candidate gene for metabolic engineering in improving the medicinal quality of A. officinarum rhizome.
Aldose-Ketose Isomerases ; genetics ; Alpinia ; chemistry ; enzymology ; genetics ; Amino Acid Sequence ; DNA, Complementary ; genetics ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Monoterpenes ; metabolism ; Phylogeny

OBJECTIVETo investigate the effect of herba schizonepetae volatile oil (STO) on the activity of 5-lipoxygenase (5-LO), so as to elucidate its mechanisms of anti-inflammatory action which is related to the arachidonic acid (AA) metabolism.

METHODThoracic cavity leukocytes from the pleurisy model rat induced by injecting 1%-carrageenan into the pleural cavity were collected. Then 0. 4 mL cell suspension including 2 x 10(7) cells per millilitre were used as the reaction system in vitro. STO in different concentrations (final concentration 0.011, 0.022, 0.043, 0.087, 0.179, 0.255, 0.364 g x L(-1)), zileuton (final concentration 0.625 x 10(-3) g x L(-1)), and DMSO in the same volume were added into the reaction tube respectively. The reaction tubes were incubated at 37 degrees C for 20 min and CaCl2 (final concentration 2 mmol x L(-1)), MgCl2 (final concentration 0.5 mmol x L(-1)), exogenous AA (final concentration 200 micromol x L(-1)) and A23187 (final concentration 5 micromol x L(-1)) were added in turns during this period. The reaction tubes were mixed and continuously incubated at 37 degrees C for 30 min. After terminating reaction by adding methanol, the metabolites of 5-LO, leukotriene B4 (LTB4) and 5-hydroxy-6, 8, 11, 14-eicosatetraenoic acid (5-HETE), were extracted, separated and detected by means of RP-HPLC.

RESULTCompared with control group, STO significantly inhibited the biosynthesis of LTB4 and 5-HETE at final concentration between 0. 022 g x L(-1) and 0.364 g x L(-1) (P < 0.05 or 0.001) in dose dependence manner, and its IC50 value was 0.124 g x L(-1) and 0.142 g x L(-1) for LTB4 and 5-HETE, respectively.

CONCLUSIONSTO can inhibited the activity of 5-LO, which is an important enzyme of AA metabolism, in rat thoracic cavity leukocytes in a dose-dependent manner in vitro. It is suggested that the mechanism of anti-inflammatory action of STO is related to its inhibiting the activity of 5-LO and decreasing the level of major inflammatory mediators LTB4.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Arachidonate 5-Lipoxygenase ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Leukocytes ; drug effects ; enzymology ; Male ; Oils, Volatile ; pharmacology ; Plant Oils ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Thoracic Cavity ; drug effects ; enzymology ; immunology