Microfluidics:Emerging prospects for anti-cancer drug screening

Cancer constitutes a heterogenic cellular system with a high level of spatio-temporal complexity.Recent discoveries by systems biologists have provided emerging evidence that cellular responses to anti-cancer modalities are stochastic in nature.To uncover the intricacies of cell-to-cell variability and its relevance to cancer therapy,new analytical screening technologies are needed.The last decade has brought forth spectacular innovations in the field of cytometry and single cell cytomics,opening new avenues for systems oncology and high-throughput real-time drug screening routines.The up-and-coming microfluidic Lab-on-a-Chip(LOC)technology and micrototal analysis systems(μTAS)are arguably the most promising platforms to address the inherent complexity of cellular systems with massive experimental parallelization and 4D analysis on a single cell level.The vast miniaturization of LOC systems and multiplexing enables innovative strategies to reduce drug screening expenditures while increasing throughput and content of information from a given sample.Small cell numbers and operational reagent volumes are sufficient for microfluidic analyzers and,as such,they enable next generation high-throughput and high-content screening of anticancer drugs on patient-derived specimens.Herein we highlight the selected advancements in this emerging field of bioengineering,and provide a snapshot of developments with relevance to anti-cancer drug screening routines.

HPLC Method Research of Picoplatin-a New Platinum Anti-cancer Drug and Its Impurities

Purpose: To establish a HPLC testing method of the content of bulk picoplatin and its impurities. Method: the separation was perform on a C18 column(4.6 mm×250 mm, 5 m) with potassium dihydrogen phosphate-aceton-itrile as the mobile phase at a flow rate of 1.0 mL/min. The detecting wavelength was set at 210 nm, and the column temperature was set at 30℃. Result: in the method validation, the linear relationship modulus of picoplatin is 0.9999, the systemic precision is 0.44%, the method precision is 0.74%, the average recovery rate is 99.62%, the LOD and LOQ of picoplatin is 0.2 ng and 1.0 ng. The average resolution of picoplatin and its impurities is more than 2. Conclusion: The established method is good specificity, high sensitivity, and good repeatability which could provide scientific evidence for the quality control of picoplatin and its impurities.

Research Progress of Radical Treatment of Helicobacter Pylori Based on Drug Sensitivity Test as First-Line Treatment

Helicobacter pylori (HP) infection is a global problem that affects about half of the world’s population and requires sufficient attention in clinical and scientific work. Due to differences in economic and medical conditions among countries around the world, there is currently no unified treatment plan for anti-HP. In China, empirical quadruple therapy is mainly used. With the abuse of antibiotics, many patients face the problem of secondary eradication after failure, and the resistance rate of HP is gradually increasing. After eradication failure, drug sensitivity cultivation is carried out to choose sensitive antibiotics for treatment. A new strategy is currently needed to address how to improve the eradication rate of HP during the first eradication. This article aims to discuss the first-line treatment plans and research progress for eradicating HP based on drug sensitivity testing before eradication. Compared with traditional empirical therapies, treatment based on drug sensitivity results can effectively improve the eradication rate of HP, and reduce drug resistance rates, and adverse reactions, among other benefits. .

Carbohydrate-associated epitope-based anti-cancer drugs and vaccines

RP215 is one of the three thousand monoclonal antibodies (Mabs) which were generated against the OC-3-VGH ovarian cancer cell line. RP215 was shown to react with a carbohydrate-associated epitope located specifically on glycoproteins, known as CA215, from cancer cells. Further molecular analysis by matrix adsorption laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) revealed that CA215 consists mainly of immunoglobulin super-family (IgSF) proteins, including immunoglobulins, T-cell receptors, and cell adhesion molecules, as well as several other unrelated proteins. Peptide mappings and glycoanalysis were performed with CA215 and revealed high-mannose and complex bisecting structures with terminal sialic acid in N-glycans. As many as ten O-glycans, which are structurally similar to those of mucins, were also identified. In addition, two additional O-linked glycans were exclusively detected in cancerous immunoglobulins but not in normal B cell-derived immunoglobulins. Immunizations of mice with purified CA215 resulted in the predominant generation of RP215-related Mabs, indicating the immunodominance of this carbohydrate-associated epitope. Anti-idiotype (anti-id) Mabs of RP215, which were generated in the rat, were shown to contain the internal images of the carbohydrate-associated epitope. Following immunizations of these anti-id Mabs in mice, the resulting anti-anti-id (Ab3) responses in mice were found to be immunologically similar to that of RP215. Judging from these observations, anti-id Mabs, which carry the internal image of the RP215-specific epitope, may be suitable candidates for anticancer vaccine development in humans.

Patch testing and cross sensitivity study of adverse cutaneous drug reactions due to anticonvulsants: A preliminary report

AIM To evaluate the utility of patch test and cross-sensitivity patterns in patients with adverse cutaneous drug reactions(ACDR) from common anticonvulsants. METHODS Twenty-four(M:F = 13:11) patients aged 18-75 years with ACDR from anticonvulsants were patch tested 3-27 mo after complete recovery using carbamazepine, phenytoin, phenobarbitone, lamotrigine, and sodium valproate in 10%, 20% and 30% conc. in pet. after informed consent. Positive reactions persisting on D3 and D4 were considered significant. RESULTS Clinical patterns were exanthematous drug rash with or without systemic involvement(DRESS) in 18(75%), Stevens-Johnsons syndrome/toxic epidermal necrolysis(SJS/TEN) overlap and TEN in 2(8.3%) patients each, SJS and lichenoid drug eruption in 1(4.2%) patient each, respectively. The implicated drugs were phenytoin in 14(58.3%), carbamazepine in 9(37.5%), phenobarbitone in 2(8.3%), and lamotrigine in 1(4.7%) patients,respectively. Twelve(50%) patients elicited positive reactions to implicated drugs; carbamazepine in 6(50%), phenytoin alone in 4(33.3%), phenobarbitone alone in 1(8.3%), and both phenytoin and phenobarbitone in 1(8.33%) patients, respectively. Cross-reactions occurred in 11(92%) patients. Six patients with carbamazepine positive patch test reaction showed cross sensitivity with phenobarbitone, sodium valproate and/or lamotrigine. Three(75%) patients among positive phenytoin patch test reactions had cross reactions with phenobarbitone, lamotrigine, and/or valproate. CONCLUSION Carbamazepine remains the commonest anticonvulsant causing ACDRs and cross-reactions with other anticonvulsants are possible. Drug patch testing appears useful in DRESS for drug imputability and cross-reactions established clinically.

Bioengineered humanized livers as better three-dimensional drug testing model system

AIM To develop appropriate humanized three-dimensional ex-vivo model system for drug testing. METHODS Bioengineered humanized livers were developed in this study using human hepatic stem cells repopulation within the acellularized liver scaffolds which mimics with the natural organ anatomy and physiology. Six cytochrome P-450 probes were used to enable efficient identification of drug metabolism in bioengineered humanized livers. The drug metabolism study in bioengineered livers was evaluated to identify the absorption, distribution, metabolism, excretion and toxicity responses.RESULTS The bioengineered humanized livers showed cellular and molecular characteristics of human livers. The bioengineered liver showed three-dimensional natural architecture with intact vasculature and extra-cellular matrix. Human hepatic cells were engrafted similar to the human liver. Drug metabolism studies provided a suitable platform alternative to available ex-vivo and in vivo models for identifying cellular and molecular dynamics of pharmacological drugs.CONCLUSION The present study paves a way towards the development of suitable humanized preclinical model systems for pharmacological testing. This approach may reduce the cost and time duration of preclinical drug testing and further overcomes on the anatomical and physiological variations in xenogeneic systems.

Control the Silver Content in the Preparation Process of Platinum Group Anti-cancer Drugs

In order to control the silver content in the preparation process of platinum group anti-cancer drugs, we put two kinds of color reagent to color in the production process of the platinum anti-cancer drugs by UV spectra measurement to control drugs production of platinum anticancer, thus we could control the silver content in the drugs so that it meets the pharmacopoeia standards of US and

Threefold Increase in the Number of Drug Resistant TB Cases after Introduction of Universal Drug Susceptibility Testing: Experiences from Two South India Districts

Background: In India, tuberculosis (TB) is a major public health problem, and the advent of drug resistance TB (DR-TB) has worsened the situation. The Revised National TB Control Programme (RNTCP) has introduced universal drug susceptibility testing (UDST) for all diagnosed TB cases in 2018. We conducted this study to know the advantage of implementing UDST when compared to selective testing existent in 2017 on key diagnostic cascade parameters and to identify the challenges in the implementation of UDST. Methods: The study was conducted in two districts of Karnataka, India during January 2017-December 2018. The quantitative part consisted of before-and-after design and the qualitative part consisted of descriptive design. Results: In 2017 (during selective testing/“before” period) out of the 2440 TB patients, 80 (3%) were diagnosed with Isoniazid and Rifampicin resistance patients;in contrast in 2018 (during UDST/“after” period) of the 5129 TB patients 258 (5%) were diagnosed with Isoniazid and Rifampicin resistance. However, the proportion of eligible patients tested for rifampicin resistance during the “after” period was 60% when compared to 100% during the “before” period and median turnaround time for testing was also longer during the “after” period when compared to the “before” period (32.5 days vs 27.5 days). Major reasons for these two gaps were found to be difficulties in collecting sputum specimens and transportation. Conclusion: The rollout of UDST has led to a three-fold increase in a number of DR-TB cases detected in the region. There is a need for the programme to increase the proportion tested for DST by increasing the laboratory capacity and address the challenges in sputum collection and transportation.

Regulation of “Checkpoint Molecule” Sheds New Light on Anti-Cancer Drug Development

The rivalry between T cells and tumor cells somewhat mimics the scene of 'Tom and Jerry,' an animated series in which Tom (a house cat) rarely succeeds in catching Jerry (a mouse), mainly because of Jerry’s cleverness and cunning abilities. In a way, tumor cells are like Jerry, in terms of their crafty and sneaky features.

Selection of Test Samples for Monitoring Veterinary Drug Residue in Bred Animals

[Objective] This study aimed to compare the residue situation of cimaterol,a kind of forbidden veterinary drug in hair, urine and flesh of pig, so as to provide theoretical basis for monitoring veterinary drug residue in bred animals. [Method]Total three different concentrations of cimaterol were administered to pigs, and the residue amounts of cimaterol in pig hair, urine and flesh were monitored at different raising stage. [Result] During the administration period, the residue amount of cimaterol was highest in urine, so urine is the suitable sample for rapid detection of cimaterol in manufacturing enterprises. Cimaterol could be accumulated in pig hair,where cimaterol was metabolized slowly. Thus, pig hair can be used as the test sample for tracing illegal use of veterinary drugs and for vivo detection. Flesh can be used as test sample for direct judgment whether cimaterol residue exceeds the relevant standard. [Conclusion] This study will provide certain theoretical basis for drug monitor in animal husbandry.

Rapid testing methods for food contaminants and toxicants

Food safety is one of the major concerns in every country regardless of the economic and social development. The frequent occurrence of food scandals in the world has led the Chinese government to implement several strategies to fortify the food supply system to a high food safety standard. This relies heavily on laboratory testing services but conventional methods for detection of food contaminants and toxicants are limited by sophisticated sample preparation procedures, long analysis time, large instruments and professional personnel to meet the increasing demands. In this review, we have incorporated most of the current and potential rapid detection methods for many notorious food contaminants and toxicants including microbial agents, toxic ions, pesticides, veterinary drugs and preservatives, as well as detection of genetically modified food genes and adulterated edible oil. Development of rapid, accurate, easy-to-use and affordable testing methods could urge food handlers and the public to actively screen for food contaminants and toxicants instead of passively relying on monitoring by the government examination facility. This review also provides several recommendations including how to encourage the public to engage in the food safety management system and provide optimal education and financial assistance that may improve the current Chinese food safety control system.

Monitoring the hydrolyzation of aspirin during the dissolution testing for aspirin delayed-release tablets with a fiber-optic dissolution system

The purpose of this study was to investigate the hydrolyzation of aspirin during the process of dissolution testing for aspirin delayed-release tablets. Hydrolysis product of salicylic acid can result in adverse effects and affect the determination of dissolution rate assaying. In this study, the technique of differential spectra was employed, which made it possible to monitor the dissolution testing in situ. The results showed that the hydrolyzation of aspirin made the percentage of salicylic acid exceed the limit of free salicylic acid （4.0）, and the hydrolyzation may affect the quality detection of aspirin delayed-release tablets.

Drug Susceptibility Test of Volatile Oil of Artemisiaargyi to Avian Pathogenic Escherichia coli

The volatile oil of Artemisia argyi was extracted by ultrasonic assisted extraction, and the extraction rate of volatile oil was 0.68%. Thevolatile oil of A. argyi was emulsified with 1% Tween-80, and drug susceptibility test was conducted with avian Escherichia coli. The results showedthat the volatile oil of A. argyi had antibacterial effect against avian E. coli, and the minimal inhibitory concentration was 50 mg/mL. Taking sixcommon antibiotics as the control, drug susceptibility test was conducted with volatile oil of A. argyi. The results showed that 10 strains of E. coliwere sensitive to the volatile oil of A. argyi, three of which had different degrees of resistance and one had the tendency of resistance.

Evaluation of Abuse Drugs and Clinical Laboratory Tests Variations in Whole Blood ＆ Urine Samples of Abusers

In this study, five hundred urine samples and five hundred blood samples of abusers were examined for the presence of alkaloid substances and abuse drugs in urines and bloods. These numbers of blood and urine samples of addicts in clinics of welfare organization, during detoxification treatment or maintenance treatment were screened for abuse drugs presence. The all of samples were tested through as a view of clinical laboratory methods. Age ranges of female patients were 35 ~ 15 and age range of males patients were 45 ~ 15. All patients filled questionnaire and satisfy forms too. First, all fresh urine and blood samples were examined to confirm presence drugs abuses, depend on their addiction and treatment, so all samples were confirmed by two tests. Then they were examined to other clinical laboratory tests. All data were analyzed by t-test and were Anova one way and two ways of Anova Turkey, and p 〈 0.05 was considered significant. The p-value of this study was p = 0.0001. The results of this study were showed that 4% of abusers had mild increase in hematocrite level and 2% of narcotic drugs abusers had mild lower level of blood sugars than normal range and 4% of participants had increase liver enzymes such ALT （alanine transferase）, AST （aspartat transferase）, ALP （alkaline phosphatease） and 1% of them had renal failure. Although blood level BUN （blood urea nitrogen） and creatinin were examined to evaluation of their renal failure .The results in Tabriz/Iran undrevision of welfare organization clinics were approximately showed that positive results of addiction are in each of urine and blood samples. Because some of abusers directly consumed full long time agonist or partial agonists＇ drugs such as methadone and buprenorphine for their maintenance therapy in clinics. Also doing test on blood samples has high importance in distinguishing and confirmation of drugs abuse in samples. Also in this study we conclude that among all drug analytical methods the cheapest and easiest test to screening opioids and other abuse drugs in urine and blood samples is strip test for rapid diagnosis, also tests on blood samples have high importance as a view point of accuracy to distinguishing of drugs abuse, and serum levels of some other parameters showed all abusers patients situation such as liver and renal dysfimction through clinical laboratory tests.

Isolation and Identification of Four Strains of Salmonella and Drug Susceptibility Test

In order to isolate and identify the pathogenic bacteria causing dead chickens in a chicken farm in Qinhuangdao area, the liver, heart and other organs of dead chickens suspected of salmonella disease were collected aseptically, and streaked on SS agar medium and chromagar medium. Transparent colonies were observed on SS agar medium, and purple and transparent colonies on CAS medium. The isolate was conducted purification, staining microscopy, biochemical tests, and 16 S rRNA sequence analysis, and the results showed that four strains of the isolated bacte-ria were salmonella. The 16 S rRNA sequence analysis of four strains of salmonella showed that the isolates shared more than 99% homology. Drug susceptibility test was performed using paper method, and the results showed that most of the strains were resistant to tilmicosin, cefradine and sul-famethoxazole, but were sensitive to ceftriaxone.

Isolation, Identification and Drug Susceptibility Test of a Strain of Escherichia coli Causing Fox Pneumonia

[Objective] The paper was to determine the pathogen causing fox pneumonia in a breeding factory in Changli County.[Method]Through autopsy, a dominant strain was isolated from the lung of dead foxes, which was then performed Gram staining, 16 S rRNA sequence analysis and biochemical identification.[Result] The strain was negative in Gram staining, and was identified as E. coli through 16 S rRNA sequence analysis and biochemical identification. Drug susceptibility test was conducted using 15 kinds of drug susceptibility papers. The E. coli was sensitive to florfenicol, enrofloxacin, ceftriaxone, norfloxacin;intermediately sensitive to amikacin, gentamicin;and strongly resistant to penicillin, ampicillin,cefradine, sulfamethoxazole, lincomycin, streptomycin and amoxicillin.[Conclusion] It is difficult to treat E. coli causing fox pneumonia with traditional antibiotics clinically.

Isolation and Identification of Pseudomonas solanacearum and Its Drug Sensitivity Test

Ten pathogenic strains were isolated fi-om gingers infected by blast, and were identified by substrate utilization test and biochemical test. The identifica- tion results showed that these ten strains accorded with the basic characteristics of Pseudomonas solanacearum. Drug sensitivity test of ten strains was carried out, and prevention agents were screened to provide an experimental basis for the control of ginger blast.

Drug Sensitivity Test and Regression Verification of Escherichia coli from Rex Rabbit

[ Objectives] The paper aimed to select drugs reasonably for treatment of rex rabbit colibacillosis, and to isolate the pathogenicity of Escherichia coll. [ Methods ] Pathogen isolation, drug sensitivity test and pathogen regression test were performed with rex rabbits killed by E. coli in clinic. [ Results] The isolate was E. coli 0-23, susceptible to amikacin and cefotaxime sodium; when the challenge dose was 1.0 mL/rabbit （about one billion E. coli）, the test animal would discharge mucous feces. [ Conclusions] The results provided model support for clinical medicine selection against rex rabbit colibacillosis.

Utilization of Malaria Diagnostic Tests and Receipt of Anti-Malarial Drugs by Febrile Patients Attending Outpatient Clinics of Health Centre IV Facilities in Mukono District, Uganda

Background: Failure to demonstrate the presence of malaria parasites prior to treatment with anti-malarial drugs remains a challenge in Uganda, often resulting into over-prescription of anti-malarial drugs to febrile patients suspected of malaria. The aim of this study was to describe the role of utilization of malaria diagnostic tests and associated factors in the receipt of anti-malarial drugs among febrile patients suspected of malaria. Methods: In a cross-sectional study design, client-exit interviews with febrile patients and key-informant interviews with purposively selected health workers were conducted at outpatient clinics of health centre IV facilities in Mukono district. Data entry and analysis were done using EpiData 3.2 and STATA 10 respectively. Data were described using frequency distributions and proportions. Chi square was used in two by two tables, odds ratios as the measure of association and an alpha level of 0.05 was used in all significance tests. Results: Out of 408 respondents, the majority were female (252, 61.8%) and a third of the samples were aged five years and below. The mean age in years was 3.3 with a standard deviation of 2.1. More than half of the respondents (359, 88%) utilized malaria diagnostic tests and about half (241, 59%) received anti-malarial drugs. There were no statistically significant differences between utilizers and non-utilizers in most characteristics except age, history of indoor residual spraying and perceived satisfaction with services at outpatient clinics. Utilizers were 75% less likely to receive anti-malarial drugs than non-utilizers after controlling for age, sex and residence (OR: 0.25, 95%CI: 0.09, 0.66). Frequent power cut-offs as well as limited knowledge on malaria treatment guidelines amongst laboratory personnel were some of the major limitations to microscopic diagnosis of malaria. Conclusion: Utilizers were 75% less likely to receive anti-malarial drugs as opposed to non-utilizers. This implies that increasing utilization of malaria diagnostic tests can reduce the problem of over-prescription of anti-malarial drugs by 75% among those tested for malaria, since anti-malarial drugs would be received by only those with a parasi- tologically-confirmed diagnosis of malaria. Policy implications: To overcome the problem of over-prescription of anti-malarial drugs, there must be a policy that ensures a consistent power supply in all public health laboratories. Training of health workers should encompass all cadres and work-shifts for laboratory personnel should be established to enhance utilization of malaria diagnostic tests especially at night.

Isolation, Identification and Drug Sensitivity Test of a Pathogenic Escherichia coli Strain from Minks with Diarrhea

[Objectives] The study aimed to identify the pathogen that causes diarrhea in minks. [Methods] Liver tissues were aseptically collected from dead minks with diarrhea. By bacterial isolation and culture,morphological observation,biochemical test and pathogenicity test,the isolated strain was identified as pathogenic E. coli. [Results]The pathogen causing diarrhea in minks was confirmed as a pathogenic E. coli strain. Drug sensitivity test indicated that the isolated pathogenic E. coli strain was highly sensitive to ceftazidime,cefotaxime,enrofloxacin,florfenicol and cephradine,moderately sensitive to ampicillin,ciprofloxacin,amikacin,doxycycline,lincomycin and gentamycin,and resistant to amoxycillin,neomycin,spectinomycin,polymyxin and penicillin. [Conclusions] This study provided reference for the prevention and control of abortion in female minks in Qinhuangdao region.