Meta-analysis of Adverse Drug Reaction of Hematological System by PARP Inhibitors / 中国药房

OBJECTIVE：To evaluate the safety of PARP inhibitors in hematological system ，and to provide evidence-based evidence for rational drug use in the clinic. METHODS ：Retrieved from PubMed ，Embase，Cochrane Library ，ScienceDirect， CNKI，CBM，VIP and Wanfang data from May 2014 to June 2019，randomized controlled trials （RCTs）about PARP inhibitors or PARP inhibitors combined with chemical treatment drugs （trial group ）versus chemical treatment drugs alone ，placebo alone or chemical treatment drugs combined with placebo （control group ）were collected. After literature screening ，data extraction and quality evaluation with bias risk assessment tool recommended by Cochrane systematic evaluator manual 5.1.0，and Meta-analysis was performed by using Rev Man 5.3 software，and sensitivity analysis and publication bias analysis. RESULTS ：A total of 10 RCTs were included ，involving 3 129 patients. Meta-analysis showed that the incidence of anemia ≥grade 3 [RR＝7.27，95％CI （2.74，19.27），P＜0.000 1]，neutropenia≥grade 3 [RR＝2.46，95％CI（1.43，4.24），P＝0.001]，and leukopenia ≥grade 3 in trial group [RR ＝1.71，95％CI（1.15，2.54），P＝0.008] in trial group were significantly higher than control group. There was no statistically significant difference in the incidence of thrombocytopenia ≥grade 3 between two groups [RR ＝3.54，95％CI（0.66， 19.05），P＝0.14]. Results of sub-group analysis showed that the incidence of an emia≥grade 3 and neutropenia ≥grade 3 in the patients receiving PARP inhibitors alone ，PARP inhibitors combined with chemical treatment dr ugs as well as the incidence of leukopenia≥grade 3 in the patients receiving PARP inhibitors （No.2018FH001-096） combined with chemical treatment drugs were significantly higher than those receiving placebo alone ，chemical treatment com drugs alone or chemical treatment drugs combined with placebo （P＜0.05）. Sensitivity analysis supported the above results howerer，publication bias was possibility. CONCLUSIONS ：PARP inhibitor in the treatment of cancer can cause hematological system adverse drug reaction ，mainly manifesting as anemia ，neutropenia and leukopenia. These results should be interpreted with caution.

Role of rate-limiting step of mevalonate pathway in improving lycopene production in Escherichia coli / 生物工程学报

The introduction of the mevalonate pathway (MVA pathway) in recombinant Escherichia coli can improve the synthesis of terpenoids. But the imbalance expression of MVA pathway genes and accumulation of intermediates inhibit cell growth and terpenoids production. In this study, each gene of MVA pathway and key genes of lycopene synthesis pathway were cloned in plasmid to express in the recombinant E. coli LYC103 with optimizing the expression of the key genes of the 2-methyl-D-erythritol-4-phosphate pathway (MEP pathway), chromosome recombinant MVA pathway and the lycopene synthesis pathway. The results showed that the overexpression of ispA, crtE, mvaK1, idi and mvaD genes did not affect the cell growth, while lycopene production increased by 13.5%, 16.5%, 17.95%, 33.7% and 61.1% respectively, indicating that these genes may be the rate-limiting steps for the synthesis of lycopene. mvaK1, mvaK2, mvaD of MVA pathway were the rate-limiting steps and were in an operon. The mvaK1, mvaK2, mvaD operon was regulated by mRS (mRNA stabilizing region) library in front of mvaK1, obtaining strain LYC104. Lycopene yield of LYC104 was doubled and cell growth was increased by 32% compared with the control strain LYC103. CRISPR-cas9 technology was used to integrate idi into chromosome at lacZ site to obtain LYC105 strain. Cell growth of LYC105 was increased by 147% and lycopene yield was increased by 2.28 times compared with that of LYC103. In this study, each gene of lycopene synthesis pathway was expressed in plasmid to certify the rate-limiting gene based on the complete MVA pathway on the chromosome. Then the rate-limiting gene was integrated in chromosome with homologous recombination to release the rate-limiting, which providing a new strategy for the construction of high-yield strains for metabolic engineering.

Integrating balanced mevalonate pathway into chromosome for improving lycopene production in Escherichia coli / 生物工程学报

Isoprenoids are all derived from two five-carbon building blocks called isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), which are synthesized either by the mevalonate (MVA) pathway or 2-C-methyld-D-erythritol-4-phosphate (MEP) pathway. In this study, the MVA pathway genes were integrated into the chromosome of LYC101, in which the expression of key genes in the MEP synthesis pathway and lycopene synthesis pathway were optimized by artificial regulatory parts, to further improve the production of isoprenoids in Escherichia coli. The plasmids pALV23 and pALV145 were screened from a plasmid library that constructed by using the RBS library to link the genes of the MVA pathway, which greatly increased the production of β-carotene. The effects of plasmids pALV23 and pALV145 on the lycopene production in low and high lycopene production strain, LYC001 and LYC101, were compared, respectively. The production of lycopene was increased by plasmids pALV23 and pALV145 in both strains. In high lycopene production strain LYC101, pALV23 produced more lycopene than pALV145. Then, the MVA gene together of promoter of pALV23 was integrated into the chromosome of LYC101 at poxB site using method of homologous recombination helped by CRISPR-Cas9 system, resulted in genetically stable strain, LYC102. The yield of lycopene of LYC102 was 40.9 mg/g DCW, 1.19-folds higher than that of LYC101, and 20% more than that of LYC101 with pALV23. Simultaneous expression of MVA pathway and MEP pathway in recombinant E. coli can effectively increase the yield of terpenoids. In this study, a plasmid-free, genetically stable, high-yielding lycopene strain was constructed, which could be used for industrialization. Also, the platform strain can be used for the synthesis of other terpenoids.

Simultaneous Determination of Carbamazepine, Venlafaxine, Rosiglitazone and Nifedipine in Human Plasma by UPLC-MS/MS / 中国药房

OBJECTIVE:To develop a method for simultaneous determination of carbamazepine,venlafaxine,rosiglitazone and nifedipine in human plasma.METHODS:UPLC-MS/MS method was adopted to determine plasma sample after precipitated with methanol (containing 0.1％ formic acid) using pioglitazone hydrochloride as internal standard.The determination was performed on Waters ACQUITY UPLC HSS C18 column with mobile phase consisted of aqueous solution containing 0.01％ formic acid-methanol containing 0.01％ formic acid (gradient elution) at the flow rate of 0.3 mL/min.The column temperature was 50 ℃,and sample size was 5 μL.ESI was used for positive ion scanning by multi reaction monitoring mode.The ion pairs for quantitative analysis were m/z 237.00 to 194.05 (carbamazepine),m/z 278.20 to 58.10 (venlafaxine),m/z 358.08 to 135.04 (rosiglitazone),m/z 347.15 to 315.17 (nifedipine),m/z 357.09 to 134.06 (internal standard).RESULTS:The liner ranges of carbamazepine,venlafaxine hydrochloride,rosiglitazone hydrochloride and nifedipine were 2.00-2 000.00 (r=0.995 9,n=5),2.12-2 120.00(r=0.990 5,n=5),2.00-2 000.00(r=0.991 5,n=5) and 2.04-1 020.00(r=0.991 0,n=5) ng/mL;the minimum detection limits were 0.200,0.106,0.100,1.020 ng/mL,respectively.RSDs of inter-day and intra-day were less than 15％.The absolute values of RE were less than 15％.The extraction recoveries were 65.66％-96.36％ (RSD％ ＜15％,n=5),and matrix effects ranged 80.99％-114.33％.The plasma concentrations of carbamazepine in 2 epileptic patients were (1 500.41 ± 169.11),(1 159.01 ±59.24) ng/mL(RSD were 11.27％,5.60％,n=3).The plasma concentrations of nifedipine in 2 hypertensive patients were (14.68 ±1.92),(21.18 ± 3.59)ng/mL (RSD were 16.98％,13.10％,n=3).CONCLUSIONS:The method is simple,specific,sensitive,accurate and suitable for the monitoring of plasma concentration and pharmacodynamic study of above drugs.

Improving β-carotene production in Escherichia coli by metabolic engineering of glycerol utilization pathway / 生物工程学报

Glycerol is a byproduct during biodiesel production. It is an important feedstock for fermentation due to its low price and high reduced status. Multiple genes of the glycerol utilization pathway were modulated in a previously engineered high β-carotene producing Escherichia coli strain CAR015 to enhance glycerol utilization capability for improving isoprenoids production. The glpR gene, encoding glycerol 3-phosphate repressor, was firstly deleted. The glpFK, glpD and tpiA genes were then modulated by three artificial regulatory parts, M1-37, M1-46 and M1-93, respectively. β-carotene titer reached 64.82 mg/L after modulating glpD with M1-46, which was 4.86 times higher than that of CAR015, and glycerol consumption rate also increased 100%. Modulating tpiA led to a little increase of β-carotene titer, whereas modulating glpFK led to a little decrease of β-carotene titer. This demonstrated that GlpD was a rate-limiting step in glycerol utilization pathway. Q-PCR of glpF, glpK, glpD and tpiA results showed that decrease the transcription level of glpF, glpK, glpD, or decrease the transcription level of tpiA could increase the cell growth and β-carotene production, probably for the decrease of methylglyoxal toxicity. Modulating glpD and tpiA genes in combination resulted in the best strain Gly003, which produced 72.45 mg/L β-carotene with a yield of 18.65 mg/g dry cell weight. The titer was 5.23 and yield 1.99 times of that of the parent strain CAR015. Our work suggested that appropriate activation of glpD and tpiA genes in glycerol utilization pathway could effectively improve β-carotene production. This strategy can be used for production of other terpenoids in E. coli.

Simultaneous Identification and Quantification of 9 Drugs of Opiates, Ketamine and Amphetamines in Human Plasma Using UPLC-MS/MS for Doping Control / 中国运动医学杂志

Objective To develop the method for simultaneous identification and quantification of morphine,6-monoactylmorphine,codeine,heroin,ketamine,3,4-Methylenedioxyamphetamine (MDA),3,4-methylenedioxymethamphetamine (MDMA),amphetamine and methylamphetamine in human plasma.Methods UPLC-MS/MS was adopted to analyze plasma with protein precipitated using 10％ trichloroacetic acid.Plasma samples were separated on ACQUITY UPLC HSS C18 column with aqueous solution (0.01％ formic acid)-methanol (0.01％ formic acid) as the mobile phase,and at a flow rate of 0.3 mL·min-1.The multiple reaction monitoring (MRM) mode was performed combined with the positive ion mode for quantification in four sections.Results The retention time,the characteristic fragment ions,and the relative abundance ratio of the molecular ion peak could be used to identify these nine compounds sensitively.The liner calibration curve of morphine,codeine,heroin,ketamine,6-monoacetylmorphine,MDA,MDMA,amphetamine and methamphetamine were obtained in the concentration range of 5.00×10-3～5.00 (r=0.9934),1.00× 10-2～ 10.00 (r=0.9905),1.00× 10-2～ 10.00 (r=0.9929),2.50× 10-3 ～2.50 (r=0.9960),5.00× 10-3 ～ 5.00 (r=0.9925),5.00× 10-4 ～ 5.00 (r=0.9910),5.00× 10-4 ～ 5.00 (r=0.9924),5.00× 10-4 ～ 5.00 (r=0.9920) and 5.00×10-4～5.00 μg·mL-1 (r=0.9900) respectively.The lowest detection limit was 1.00,1.00,1.00,0.50,0.50,0.10,0.10,0.10 and 0.10 ng· mL-1 respectively.The relative standard deviation (RSD) of the inter-day and intra-day were less than 18％.The relative recovery was more than 60％,and the RSD were less than 15％.Conclusion The method is accurate,sensitive and suitable for identification and quantification of 9 drugs of opiates,ketamine and amphetamines in human plasma.

Blood testosterone levels of patients with mumps orchitis / 中国基层医药

Objective To analyze the effects of the mumps virus on testicular function and evaluate the value of blood testosterone in acute orchitis condition and prognosis by observing the testosterone levels of peripheral blood in the mumps orchitis patients.Methods Thirty patients with acute mumps orchitis (mumps orchitis group)and 28 patients with mumps only without any major complications (mumps group)were enrolled in the study.At the same time,we selected 20 healthy males in our hospital as healthy controls.All cases were treated by ribavirin (10mg/kg). The testosterone was tested by chemiluminescence method,in 0,7,14 days respectively.The variance analysis and LSD -t test were used to compare differences of blood testosterone in each group,and Pearson method was used to analyze the correlation of blood testosterone and testicular swelling day times,blood amylase,level of neutrophils,IL -6 and CRP.Results The testosterone of acute orchitis [(1.51 ±0.26)ng/mL]was lower than mumps group and healthy controls.The difference was significant(F =99.36,P <0.01).The level of blood testosterone was significantly increased in the treatment for 14 days (t =13.03,P <0.01),and there was no significant difference compared with healthy controls (t =1.23,P =1.23).Pearson correlation analysis found that blood testosterone levels were negatively correlated with the days of testicular swelling,the levels of IL -6 and CRP(r =0.678,P <0.01 and r =0.528,P =0.000;r =0.442,P =0.000).Conclusion This study showed that MuV infection inhibited testosterone synthesis in Leydig cells.In other words,the level of blood testosterone is related to the severity of the orchitis,which can be widely used in clinical.

Production of coenzyme Q10 by metabolically engineered Escherichia coli / 生物工程学报

Coenzyme Q10 (CoQ10) is a lipophilic antioxidant that improves human immunity, delays senility and enhances the vitality of the human body and has wide applications in pharmaceutical and cosmetic industries. Microbial fermentation is a sustainable way to produce CoQ10, and attracts increased interest. In this work, the native CoQ8 synthetic pathway of Escherichia coli was replaced by the CoQ10 synthetic pathway through integrating decaprenyl diphosphate synthase gene (dps) from Rhodobacter sphaeroides into chromosome of E. coli ATCC 8739, followed by deletion of the native octaprenyl diphosphate synthase gene (ispB). The resulting strain GD-14 produced 0.68 mg/L CoQ10 with a yield of 0.54 mg/g DCW. Modulation of dxs and idi genes of the MEP pathway and ubiCA genes in combination led to 2.46-fold increase of CoQ10 production (from 0.54 to 1.87 mg/g DCW). Recruiting glucose facilitator protein of Zymomonas mobilis to replace the native phosphoenolpyruvate: carbohydrate phosphotransferase systems (PTS) further led to a 16% increase of CoQ10 yield. Finally, fed-batch fermentation of the best strain GD-51 was performed, which produced 433 mg/L CoQ10 with a yield of 11.7 mg/g DCW. To the best of our knowledge, this was the highest CoQ10 titer and yield obtained for engineered E. coli.

Expression levels of IL-37 and HBeAg seroconversion in chronic HBV patients / 中国免疫学杂志

Objective:Detecting serum IL-37 concentrations and HBeAg seroconversion in chronic hepatitis B virus(HBV) patients during the treatment of telbivudine( LDT).Methods:Fifty patients with chronic hepatitis B were included in the study;and 20 healthy people were selected as the control group.The expression levels of interleukin ( IL)-37, IL-6 and CRP were measured by enzyme-linked immunosorbent assay.As the same time,the relationship between serum IL-37 concentrations and HBeAg seroconversion was dynamically observed at 0 w,12 w,24 w,36 w,48 w after treatment.Results:The serum levels of IL-37 in chronic HBV patients were higher than the control group at baseline(q=22.716,P0.05).Conclusion:IL-37 may be involved in the process of HBV infection in CHB patients,and can reflect the degree of liver’ s inflammatory damage.IL-37 may play a significant role in the immune response of CHB patients with HBeAg seroconversion.In future,IL-37 may be an effective therapeutic measure to HBV infection.

Expression of Th17, CD4 +CD25 +Treg, HLA-DR mRNA in peripheral blood of children with hand, foot and mouth diseases / 中华临床感染病杂志

Objective To investigate the expressions of Th17, CD4 +CD25 +Treg, HLA-DR mRNA in peripheral blood of children with EV71-induced hand, foot and mouth diseases ( HFMD ) and their clinical significance.Methods Stratified random sampling was used to select 60 children with HFMDs from Liaocheng People’s Hospital from February to October, 2014, including 20 mild, 20 severe and 20 critically ill cases.Twenty healthy children were also enrolled as the control group.All the children with HFMDs were given ribavirin (10 mg/kg) for the treatment.The percentages of Th17 and CD4 +CD25 +Treg cells in CD4 +T cells of peripheral blood were determined by flow cytometry, and the expression of HLA-DR mRNA in peripheral blood mononuclear cells was detected by reverse transcription polymerase chain reaction ( RT-PCR).Analysis of variance and SNK-q test were used to compare the expressions of Th17, CD4 +CD25 +Treg and HLA-DR mRNA among groups, and Pearson correlation analysis was performed to reveal the correlations between HLA-DR mRNA and Th17, CD4 +CD25 +Treg.Results Compared with the control group, the expression of Th17 was increased, while CD4 +CD25+Treg and HLA-DR mRNA expressions were decreased in children with HFMDs on d1 of treatment (F=310.4, 81.5 and 545.4, P0.05), but Th17 level in fatal was still higher and CD4 +CD25 +Treg level was still lower than those in control group (t=16.4 and 12.0, P<0.05).After 10 d of treatment, HLA-DR mRNA expressions in mild group and severe group were increased to the normal level.HLA-DR mRNA expression in surviving patients of critical ill group was still lower than that in mild group and severe group (P<0.05), but was higher than that in fatal patients (t=7.8, P<0.05).Pearson correlation analysis showed that, HLA-DR mRNA was negatively correlated with Th17 level (r=-0.770, P<0.01), and positively correlated with CD4 +CD25 +Treg level (r=0.883, P<0.01).Conclusion The expressions of Th17, CD4 +CD25 +Treg cells, and HLA-DR mRNA are correlated with the severity of HFMD, and may be used for evaluation of disease severity and prediction of disease outcomes.

Modulating expression of key genes within β-carotene synthetic pathway in recombinant Escherichia coli with RBS library to improve β-carotene production / 生物工程学报

β-carotene belongs to carotenoids family, widely applied in pharmaceuticals, neutraceuticals, cosmetics and food industries. In this study, three key genes (dxs, idi, and crt operon) within β-carotene synthetic pathway in recombinant Escherichia coli strain CAR005 were modulated with RBS Library to improve β-carotene production. There were 7%, 11% and 17% increase of β-carotene yield respectively after modulating dxs, idi and crt operon genes with RBS Library, demonstrating that modulating gene expression with regulatory parts libraries would have more opportunities to obtain optimal production of target compound. Combined modulation of crt operon, dxs and idi genes led to 35% increase of β-carotene yield compared to parent strain CAR005. The optimal gene expression strength identified in single gene modulation would not be the optimal strength when used in combined modulation. Our study provides a new strategy for improving production of target compound through modulation of gene expression.

Effect of bundle hand hygiene intervention in controlling healthcare-asso-ciated infection in a primary comprehensive hospital / 中国感染控制杂志

Objective To explore the effect of bundle hand hygiene intervention in controlling healthcare-associated infec-tion(HAI)in a primary comprehensive hospital,so as to improve hand hygiene compliance and correct rate,and reduce AHI rate. Methods In January-June 2014,bundle hand hygiene intervention among health care workers (HCWs)in a hospital was performed,hand hygiene knowledge awareness rate,hand hygiene compliance and correct rate,hand sanitizer usage and HAI rate before and after intervention were compared. Results After performing intervention for six months,the awareness rate of hand hygiene knowledge(concept,significance,indications,methods,sanitizer use)of HCWs improved com-pared with before intervention (P<0.05);hand hygiene compliance and correct rate were significantly higher than before inter-vention (77.92% vs 49.78% ;76.47% vs 37.72% )(P<0.05). Hand sanitizer usage increased from 2.14mL/bed-day to 4.63 mL/bed-day ,HAI rate decreased from 1.97% to 1.54% (P<0.05).Conclusion Bundle hand hygiene intervention can improve HCWs’knowledge awareness,compliance and execution rate of hand hygiene,and effectively reduce HAI rate.

Influence of beryllium ions on genotype of Porphyromonas gingivalis fimbriae gene cluster / 中国组织工程研究

BACKGROUND:In oral warm and moisture circumstance, al oy which contains Be is easily to be eroded to release Be2+. But there is stil no research focusing on beryl ium influence on genotype of Porphyromonas gingivalis fimbriae gene cluster. OBJECTIVE:To investigate Be 2+effect on genotype of Porphyromonas gingivalis fimbriae gene cluster. METHODS:The revived Porphyromonas gingivalis was resuscitated for 48 hours in the anaerobic culture medium with different concentration of Be 2+(10×10-6, 5×10-6, 1.25×10-6). Through PCR amplification and sequencing, we investigated the effects of Be2+RESULTS AND CONCLUSION:(1) When Be on genotypes of Porphyromonas gingivalis fimbriae gene cluster. 2+concentration was 5×10-6, we found the peak of 217 and 257 sites on DNA sequence expressing G/A overlap peak, different from G single peak of the other groups, suggesting the suspicious bases changes, part of the single base G mutated into A. (2) On al concentrations, we found a base group composed of seven A bases was inserted into the 101 site of DNA sequence. Up to now, there is no direct contacts of the mutations occurring to Be2+concentration. Changes of gene may lead to the shifting of the reading frame, the abnormal synthesis of proteins, the change of Porphyromonas gingivalis fimA gene toxicity, and lastly the unbalance of the micro-ecological environment.

Modulation of isoprenoid gene expression with multiple regulatory parts for improved beta-carotene production / 生物工程学报

Strong promoters might not be optimal to obtain maximum metabolic flux towards desired products, whereas modulating gene expression with multiple regulatory parts is an option to obtain optimal expression strength. Therefore, we assessed the difference of impact on beta-carotene production between modulating isoprenoid gene expression with multiple regulatory parts and strong promoter, to improve beta-carotene production through combined modulation of essential isoprenoid genes. Eight isoprenoid genes were modulated with six artificial regulatory parts having a wide range of strengths to assess their effects on beta-carotene production. Optimal strength for each isoprenoid gene expression was identified, leading to 1.2 to 3.5-fold increase in beta-carotene production. In contrast to previous reports, our work suggests that modulating dxr, ispG and ispH genes with appropriate strengths increase beta-carotene production. Beta-carotene yield reached 17.59 mg/g after combined modulation of dxs and idi genes, 8-fold higher than that of the parent strain. Modulating gene expression with multiple regulatory parts was better than strong promoter, providing a new gene modulation strategy for targeted biosynthesis.