Validation of a fast and reliable liquid chromatography tandem mass spectrometry for simultaneous quantitation of Voriconazole, Posaconazole and Linezolid in human serum / 中华检验医学杂志

Objective:To establish a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of voliconazole (VRC), posaconazole (PCZ), and linazolam (LNZ) in human serum.Methods:This study is a methodological validation by LC-MS/MS. The blood concentration results of VRC, PCZ, and LNZ in our hospital′s anti-infection patients were collected. Voriconazole, Posaconazole, and Linezolid were accurately weighed and prepared. Linezolid-[2H3] was used as the internal standard. After gradient elution on the ACE PFP column, the residuals were analyzed by LC-MS/MS in the positive electrospray ionization mode and multiple reaction monitor (MRM) mode. The method′s linearity, precision, lower limit of detection, and recovery rate were validated according to standard guidelines.Results:The linear correlation coefficient ( r) of the standard curve was above 0.99 ( r>0.99). The linear range of VRC and PCZ were 0.10 mg/L~10.00 mg/L, and the lower limit of detection were 0.01 mg/L. The linear range of LNZ was 0.50 mg/L~50.00 mg/L, and the lower limit of detection was 0.05 mg/L. The recoveries of VRC, PCZ and LNZ were 90.96%-103.18%, 91.84%-99.17%, and 97.04%-100.41%, respectively. Intra-and inter-batch precision (% CV) for VRC were less than 8.30%. Intra-and inter-batch precision (% CV) for PCZ was less than 9.78%. Intra-and inter-batch precision (% CV) for LNZ was less than 7.14%. Drug concentrations in 155 cases of VRC, 44 cases of PCZ, and 59 cases of LNZ were detected. Conclusion:We have established an LC-MS/MS method for the rapid, accurate, highly specific determination of VRC, PCZ, and LNZ concentrations in human serum. This method is suitable for analyzing large clinical sample sets.

Efficacy analysis of subcutaneous injection of granulocyte-macrophage colony-stimulating factor for prevention of invasive fungal disease in patients with multiple myeloma / 白血病·淋巴瘤

Objective:To explore the efficacy of subcutaneous injection of granulocyte-macrophage colony-stimulating factor (GM-CSF) in preventing invasive fungal disease (IFD) in patients with multiple myeloma (MM).Methods:The clinical data of 222 patients who were admitted to the Second Hospital of Harbin Medical University from January 2015 to June 2021 were retrospectively analyzed. The patients was given GM-CSF (3-5 μg·kg -1·d -1, GM-CSF group) or granulocyte colony-stimulating factor (G-CSF, 2-5 μg·kg -1·d -1, G-CSF group) when neutrophils (ANC) ≤1.5×10 9/L after induction chemotherapy. Patients were discontinued when white blood cell count (WBC) ≥10.0×10 9/L. The incidence of IFD (including confirmed, clinical and proposed diagnosis) and breakthrough invasive fungal infections was compared between the two groups. Results:The incidence of IFD was 8.1% (18/222) in all patients. The incidence of IFD was 3.5% (3/85) and 10.9% (15/137) in the GM-CSF and G-CSF groups, respectively, and the difference between the two groups was statistically significant ( χ2 = 3.88, P = 0.049). In 9 patients of GM-CSF group receiving fungal infection prophylaxis and in 15 patients of G-CSF group receiving fungal infection prophylaxis, the incidence of breakthrough invasive fungal infections was 0 and 7 cases, respectively, and the difference between the two groups was statistically significant ( P = 0.022). Conclusions:GM-CSF application in MM patients can reduce the incidence of IFD and breakthrough invasive fungal infections.

Analysis on effect of antimicrobial stewardship based on information technology for 6 years / 中华急诊医学杂志

Objective:To evaluate the effectiveness of antimicrobial stewardship based on self-developed antibiotic clinical decision support system (aCDSS) in the inpatients at a tertiary hospital for consecutive 6 years, and to provide reference for rational use and antimicrobial stewardship.Methods:aCDSS was self-designed based on information technology and applied in clinical use in our hospital from 2015. Data of inpatient information and antibacterial use from January 2015 to December 2020 were collected from HIS and aCDSS. A retrospective study was conducted in all inpatients on the utilization rate and antibiotic use density.Results:Since 2015, with the comprehensive implementation of antimicrobial stewardship based on the aCDSS,there was a significant decline on the annual rate of antibiotic usage from 44.18% in 2015 to 38.70% in 2020, as well as on the usage rate of extended-spectrum antimicrobial agents including carbapenems, broad-spectrum β-lactam/β-lactamase inhibitors, tigecycline, broad-spectrum cephalosporins, fluoroquinolones, as well as glycopeptide and antifungal drugs. Compared with 2015, the usage of carbapenems, tigecycline and broad-spectrum β-lactam/β-lactamase inhibitors was declined nearly 50% in 2020, and the density of carbapenems and tigecycline were decreased by 29.6% and 7.1%, respectively in 2020. On the other side, the utilization rate and use density of narrow-spectrum cephalosporins continued to increase by year, the use density of narrow-spectrum cephalosporins accounting for 28.2% of all antibiotics in 2020.Conclusions:With the comprehensive implementation of aCDSS, the utilization rate and density of broad-spectrum and high-priced antibacterial drugs in our hospital have decreased continuously to decline in the past 6 years, while the proportion of narrow-spectrum antimicrobials has increased year by year, indicating that the structure of antimicrobial use has been continuously optimized and that antimicrobial stewardship based on the information technology have achieved remarkable results.

Effectiveness of the special-grade antimicrobial stewardship based on information technology / 中华临床感染病杂志

Objective:To evaluate the effectiveness of the information management system on the clinical application of special-grade antimicrobial.Methods:Using the established knowledge database, a computer program was designed and developed, which was embedded in the electronic medical record to intervene the clinical use of the special-grade antimicrobial since 2015. The basic information of all discharged patients from the Second Affiliated Hospital of Zhejiang University School of Medicine from January 2013 to December 2020 were extracted from the HIS system, including the medical orders for antibiotics and the drug storehouse dispensing data.The trend analysis was carried out on the changes of the use rates and antibiotic use density (AUD) of the special-grade antimicrobials in the whole hospital and intensive care units (ICU). The data were processed and analyzed using SPSS 24.0.Results:From 2013 to 2015, except for meropenem and amphotericin B, the usage rate of all special-grade antimicrobials in the whole hospital showed an upward trend ( P<0.05). The proportion of special-grade antimicrobials used in the hospital increased year by year ( χ2=7 804.081, P<0.01). The total usage rate of special-grade antimicrobials in ICU showed an upward trend year by year ( χ2=67.028, P<0.01). Since the implementation of the special-grade antimicrobial information management system in 2015, the total use rate of special-grade antimicrobials in the hospital, the use rate of various antibiotics except linezolid, amphotericin B and posaconazole, and the proportion of special-grade antimicrobials used in the hospital have all shown a downward trend year by year ( P<0.01). The total usage rate and total AUD of special-grade antimicrobials in ICU showed a decreasing trend year by year ( χ2=343.514, P<0.01, β=-0.963, P=0.002). Conclusion:The information management system for special-grade antimicrobial can effectively reduce the utilization rate and AUD of most special-grade antibiotics in hospitalized patients including ICU, and has a good clinical application value in antimicrobial stewardship.

Medication analysis and pharmaceutical care for a Parkinson′s disease patient with mental disorders / 药学实践杂志

Objective To investigate the role of clinical pharmacists in the treatment of a Parkinson′s disease patient with mental disorders.Methods Clinical pharmacists provided appropriate pharmaceutical care and optimized the treatment program based on patient′s symptom, medication history, drug interactions,and adverse drug reactions etc.Results Clinical pharmacists improved rational drug use by participating in the development of patient′s treatment program, giving patient with proper medication instruction and discharge education.Conclusion With their professional knowledge, clinical pharmacists play an important role in rational drug use and helping physicians to optimize the medication regimen.

Neuronal mitochondria and apoptosis signaling pathways play an important role in cell death during transient cerebral ischemia / 中国组织工程研究

BACKGROUND:Although the mechanism why neuronal cels wil die after transient cerebral ischemia has not been completely elucidated, many researches nowadays have investigated the pathological mechanism in the level of celular organs, such as mitochondria. OBJECTIVE:To summarize and discuss the functions of neuronal mitochondria and apoptosis signaling pathways in transient cerebral ischemia. METHODS: A computer-based online retrieval was performed to search papers in CNKI and PubMed databases using the key words of “cerebral ischemia, mitochondrion, apoptosis, reactive oxygen species, reperfusion, superoxide dismutase, nitric oxide synthase, Bcl-2 protein family, review” in Chinese and English, respectively. Papers published recently or in the prestigious journals were selected in the same field. After excluding objective-independent papers and repeated studies, 50 papers were included for further analysis. RESULTS AND CONCLUSION:Recently mitochondria are found to play an important role after transient cerebral ischemia by producing a lot of reactive oxygen species to activate many kinds of signaling pathways and regulate mitochondria-mediated apoptosis. Reactive oxygen cannot only induce biomacromolecule injury but also induce apoptosis signal transduction. Deeply investigation is needed on the pathological mechanism after transient cerebral ischemia.

Hydrogen sulfide and mild hypothermia can selectively activate synaptic NMDARs and trigger the CREB signaling pathway / 医学研究生学报

Objective Research has indicated that hydrogen sulfide(H2S) can regulate the function of N-methyl-D-aspartate re-ceptors(NMDARs) in the brain, but its effect on brain resuscitation requires further investigation.The study was to speculate the effect of H2 S on brain resuscitation as well as the underlying mechanism of neuroresuscitation by investigating the effects of hydrogen sulfide and hypo-thermia on the expression of NR2A, NR2B and phospho-cAMP response element binding protein (p-CREB) of NMDARs in the hippocampus after global cerebral ischemia following by reperfusion. Methods 100 male SD rats were randomly divided into five groups(n=20):sham operation group, model group, mild hypothermia group, NaHS group, NaHS combined mild hypothermia group.Pulsinelli-Brierley four-ves-sel occlusion method was induced to build the injury rat model by reperfusion after global cerebral ischemia .After 15 minutes'ischemia, im-mediate injection of 14μmol/kg NaHS was performed intraperitoneally on NaHS group and NaHS combined mild hypothermia group , while skin cooling(rectal temperature=32-33℃) was done on mild hypothermia group and NaHS combined mild hypothermia group .6 hours late,r hip-pocampus were extracted from rat heads.Respectively, spectrophotometer was applied to measure the content of H2S, Western blot for the expres-sions of NR2 A,NR2 B and pC-REB, and RTP-CR for mRNA level of brain derived neurotrophic (BDNF). HE staining was also performed on brain tissues 72hours after reperfusion on 4 rats from each group to evaluate the pathological changes of pyramidal neurons in CA1 region. R esul ts The content of H 2 S increased in each of the four groups after ischemia-reperfusion compared with sham operation group ( 15.2 ±2.0 nmol/g) (P0.05).The gray values of NR2A and NR2B in each group increased compared with sham operation group(P1 in NaHS group and NaHS combined mild hy-pothermia group.Compared with the expression of p-CREB(0.55 ±0.06) in model group, there were significant increases in mild hypother-mia group(0.99 ±0.15), NaHS group(1.05 ±0.12), NaHS combined mild hypothermia group(1.02 ±0.15)(P<0.05).Compared with the expression of BNDF mRNA(0.83 ±0.12) in model group, there were significant increases in mild hypothermia group (1.11 ±0.13), NaHS group(1.27 ±0.16), NaHS combined mild hypothermia group(1.35 ±0.16)(P<0.05).In comparison to model group, there were signifi-cant alleviation in the injury of pyramidal neurons in hippocampal CA1 region in mild hypothermia group, NaHS group, NaHS combined mild hypothermia group, with the best effect in NaHS combined mild hypothermia group . Conclusion Hydrogen sulfide combined mild hypo-thermia can selectively activate synaptic NMDA receptors and trigger the prosurvival CREB signaling pathway to exert brain resuscitation .

Aminoguanidine suppresses methylglyoxal-mediated oxygen-glucose deprivation injury in human brain microvascular endothelial cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the effects of aminoguanidine on methylglyoxal-mediated oxygen-glucose deprivation (OGD) injury in the cultured human brain microvascular endothelial cells (HBMEC).</p><p><b>METHODS</b>Cultured HBMEC cells were pretreated with methylglyoxal before oxygen-glucose deprivation injury. Cell vitality was determined by MTT method, cell mortality was assessed by LDH release method, cell apoptosis was examined by Annexin V/PI formation method, and the advanced glycation end products (AGEs) were detected by Western-blot.</p><p><b>RESULTS</b>Methylglyoxal induced HBMEC injury in a dose-dependent manner. At 2 mmol/L of methylglyoxal, the cell viability was 56.1% when methylglyoxal-pretreated cells exposed to oxygen-glucose deprivation, the cell inhibition rate was 90.0%. Aminoguanidine (1 mmol/L) inhibited methylglyoxal and OGD induced LDH release and Annexin V/PI formation. Furthermore, aminoguanidine (1 mmol/L) also decreased advanced glycation end products (AGEs) formation induced by methylglyoxal and oxygen-glucose deprivation.</p><p><b>CONCLUSION</b>Aminoguanidine protected methylglyoxal mediated-oxygen-glucose deprivation injury in the cultured HBMEC, which may be associated with anti-glycation activity.</p>

Identification of a HEK-293 cell line containing stably-transfected H3R gene and screening for novel non-imidazole histamine H3 receptor antagonists / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To identify a HEK293 cell line containing stably-transfected H3R gene, and to screen the novel non-imidazole compounds with H3R antagonist activity.</p><p><b>METHODS</b>The expression of rat H3 receptor in cell line was detected by RT-PCR and Western blot. An elevation of intercellular cAMP concentration induced by forskolin was measured as the index for screening compounds with H3R antagonist activity.</p><p><b>RESULTS</b>The H3R-transfected HEK-293 cells stably expressed high level of rat H3 receptor mRNA and protein. Forskolin significantly increased intercellular cAMP concentration in the H3R-transfected HEK-293 cells. H3R agonist (R)-α-methylhistamine inhibited the forskolin-induced production of intercellular cAMP. H3R antagonist thioperamide and newly synthesized non-imidazole compounds XHA23 and XHA25 blocked (R)-α- methylhistamine reversal of forskolin-induced cAMP formation in a concentration-dependent manner, and the IC50 values were 3.62 μmol/L, 0.49 μmol/L, 0.14 μmol/L, respectively.</p><p><b>CONCLUSION</b>The H3R-transfected HEK293 cells stably express high level of rat H3 receptor, and can be used for screening compounds with H3R antagonist activity. The non-imidazole compounds XHA23 and XHA25 may have H3R antagonist activity.</p>

Allogeneic hematopoietic stem cell transplantation using reduced-intensity Bu/Cy conditioning for patients with hematologic malignancies / 中国实用内科杂志

Objective To explore the efficacy of allogeneic hematopoietic stem cell transplantation using reduced-intensity Bu/Cy conditioning for patients with hematologic malignancies.Methods Five patients with hematologic malignancies were treated by allogeneic hematopoietic stem cell transplantation using reduced-intensity Bu/Cy conditioning,which consisted of busufan3~4 mg/(kg?d) for 3 days,cyclophosphamide50 mg/(kg?d) for 2 days, Ara-C 2 g/(m~2?d) for 1 or 2 days,and CsA 3 mg/(kg?d) and mycophenolate mofetil 1 g/d 7 days before the transplantation.Results Five patients established successful engraftment and no severe complications occured.After a follow-up of median 10.5(3-22)months,five patients still survived without diseases.Conclusion Reduced-intensity Bu/Cy conditioning may reduce transplantation-related toxicities.Allogeneic hematopoietic stem cell transplantation using reduced-intensity conditioning is a safe and effective option for the patients with hematologic malignancies.