The research and development of monoclonal antibodies (mAbs) is a rapidly developing field. From the first generation of murine mAbs to the fourth generation of fully human mAbs, the efficacy and safety of mAbs in the treatment of various diseases have been continuously improved. In order to regulate the development and evaluation of mAbs, drug regulatory agencies and pharmacopeias of America and China have tried to issue feasible test procedures and acceptance criteria for quality evaluation of mAbs and biosimilars. Mass spectrometry (MS) technique with high sensitivity, resolution, selectivity, and specificity has become an important tool to evaluate the quality characteristics of monoclonal antibody-related products or specify mAb quality. The research of MS-based monoclonal antibody study involves structure characterization, impurity analysis, pharmacokinetics/pharmacodynamics (PK/PD), etc. This review focuses on the current quality control requirements of mAb related products and the development of MS technique for mAb quality characterization and specification. It is expected to provide information and references for evaluating the quality of monoclonal antibodies under research and development.

Objective To observe the therapeutic effect of stellate ganglion block (SGB) combined with Tianshu capsule on migraine.Methods Ninty patients with migraine were randomly divided into the trial group (n=44, treated by SGB combined with Tianshu capsule) and control group (n=46, treated by SGB only). All patients of two groups were evaluated with numeric rating score (NRS) for pain before treatment and 1 day, 3 days, 1 week, 1 month and 3 months after treatment, and the therapeutic effects of two groups were compared.Results The NRS scores of patients in the two groups at each time point decreased, but scores of patients in the trial group were significantly lower than that in the control group at 3 days, 1 month and 3 months after treatment ( P<0.05). The effective rate of the trial group was superior to that of the control group ( P<0.05).Conclusion SGB combined with Tianshu capsule is more effective on migraine and has less side effect.

In recent years, the biopharmaceutical industry has grown rapidly, and the market size of monoclonal antibody drugs has increased significantly. Accurate structural characterization and quality control are the supporting technologies for the development of monoclonal antibody drugs. As a significant post-translational modification of antibody drugs, glycosylation has an important influence on its efficacy, stability, and immunogenicity. The existing literature usually uses liquid chromatography-mass spectrometry to perform major glycosylation modifications of monoclonal antibody drugs. Characterization, there are few studies on low-abundance glycosylation, but the characterization and control of low-abundance glycosylation cannot be ignored. In this study, we have established a qualitative and quantitative analysis technology for N-glycans based on RapiFluor-MS reagent-labeled monoclonal antibody drugs. This method has a short sample processing time and high sensitivity. It can not only characterize the main glycoforms of three monoclonal antibody drugs (adalimumab, bevacizumab, and trastuzumab) but also can quantify low-abundance N-glycans. The results of the study showed that the main glycoforms specified in the Pharmacopoeia could be detected in different batches of monoclonal antibody drugs, but the content of N-glycans in different batches of samples is not identical. After that, we analyzed the N-glycans connection sites and glycoforms at the intact glycopeptide level, further enriching the N-glycans structure information of the monoclonal antibody. The qualitative and quantitative analysis technology of N-glycans based on RapiFluor-MS reagent-labeled monoclonal antibody drugs can realize the in-depth characterization and control of glycosylation modification of monoclonal antibody drugs.

Objective To find out the satisfaction changes among young medical workers and explore the influence of the healthcare reform on such people.Methods A survey was conducted by means of random sampling among 716 medical workers under 45 years old from 6 hospitals,and combined with the survey data in 2009,young medical workers′satisfaction changes and the influencing factors were analyzed.Results Young medical workers′satisfaction is generally low,with declining satisfaction of social respect as well;satisfaction of medical workers in community medical institutions and secondary medical institutions was found declining dramatically.Conclusions Healthcare reform should pay more attention to these young people in terms of their own satisfaction,especially for those working at primary healthcare institutions.

Purpose To investigate the value of contrast enhanced ultrasound (CEUS) in evaluating blunt hepatic trauma and to provide experimental reference. Materials and Methods Fifteen healthy New Zealand white rabbits were randomly divided into groups A, B and C, with 5 rabbits in each group. Hepatic area of the rabbits in each group was stroked using a self-made mini striker with forces of 127.8 N, 213.0 N and 298.2 N, respectively, to establish rabbit models of blunt hepatic trauma. Gray-scale ultrasound (US) and CEUS were performed within 20 minutes before and after trauma. Results Hepatic capsular rupture was observed in 10 rabbits on US. Ill-defined hypoechoic areas in various degrees were observed. CEUS and time-signal intensity curve analysis showed that the normal hepatic tissue began to enhance after contrast agent injection, well-defined against non-enhanced hepatic lesions. The sensitivity of US and CEUS for blunt hepatic trauma were 66.7% (10/15) and 100.0% (15/15), respectively (P<0.05). The agreement with specimen was 40.0% (6/15) for US and 86.7% (13/15) for CEUS, respectively (P<0.05), with Kappa=0.815 for CEUS (P<0.001). Conclusion CEUS improves diagnostic accuracy of blunt hepatic trauma and accurately assesses the severity, which shows good agreement with gross specimen.

PurposeAcute lung injury/acute respiratory distress syndrome is a common severe disease, this paper aims to explore the pathological basis of diffuse comet-tail artifacts in rabbits with acute lung injury and evaluate the imaging value on pathological changes.Materials and Methods The bilateral anterior chest walls of the rabbits were divided into four zones, namely, left upper zone, right upper zone, left lower zone and right lower zone. The normal chest ultrasound images of the 6 rabbits were obtained. After oleic acid (0.1 ml/kg) was injected, the images of comet-tail artifact in the four lung zones were dynamically observed and stored for 1 hour. The chest ultrasound images were evaluated according to the ultrasound scores. The degree of pathological damage in each lung zone was assessed and analyzed in terms of the correlation with the ultrasound results.Results The animal model of acute lung injury with diffuse comet-tail artifacts was successfully made in all of the 6 rabbits. The main pathological changes of the animal model of oleic acid were diffuse pulmonary edema, congestion and congestive hemorrhage; necrosis was found in 4 rabbits. The area of pulmonary edema, congestion, and congestive hemorrhage increased gradually from top to bottom, and double diaphragm leaves were typical. A significant correlation was found between the ultrasound scores of comet-tail artifacts and the degrees of pathological damage (r=0.819,P<0.05).Conclusion The histological changes of chest diffuse comet-tail artifacts due to acute lung injury are pulmonary edema, congestion and congestive hemorrhage. The scope and density of diffuse comet-tail artifacts may indicate the degree of pulmonary parenchyma injury, which is important as reference to guide the treatment and prognosis.

To synthesize salbutamol immunogen and develop an enzyme immunoassay (ELISA), a new salbutamol immunogen was synthesized using 4-aminobenzoic acid as a linker to connect hapten with carrier protein. An enzyme immunoassay based on the antibody prepared was developed and applied to detect salbutamol residue spiked in swine liver. An unusual coating antigen, clenbuterol-ovalbumin (OVA) conjugate instead of salbutamol-OVA conjugate, was used in the immunoassay and the results were discussed based on the structures of related compounds. The antibodies showed high sensitivity in the heterologous assay when using clenbuterol-OVA as a coating antigen, with an IC50 value of 8.97 ng mL(-1) toward salbutamol. The antibodies prepared showed high cross-reactivity with clenbuterol (107%) and were promising for the simultaneous determination of salbutamol and clenbuterol residues in food and food products. Recovery rates from the salbutamol-spiked swine liver samples were in the range of 70%-99%, while the intra-assay and inter-assay coefficients of variation were <13.3% and <14.3%, respectively. In summary, the antibodies of salbutamol have been successfully prepared. Sensitive and stable analysis for the detection of salbutamol residues in swine liver was obtained based on the competitive ELISA methods developed in this study.
4-Aminobenzoic Acid ; chemistry ; Adrenergic beta-Agonists ; analysis ; immunology ; Albuterol ; analysis ; immunology ; Animals ; Antibodies ; immunology ; Antibody Specificity ; Clenbuterol ; analysis ; immunology ; Drug Residues ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Food Contamination ; Haptens ; immunology ; Immunization ; Liver ; chemistry ; Male ; Ovalbumin ; chemistry ; immunology ; Rabbits ; Serum Albumin, Bovine ; chemistry ; immunology ; Swine

In order to investigate the prevalence and track genetic and antigenic evolutions of infectious bronchitis virus (IBV) and their prevalence in Guangxi, China since 1985, gene amplification and sequencing and virus neutralization (VN) test on chicken embryo tracheal organ cultures were used in genotyping and serotyping of 28 IBV isolates during 2009-2011 in Guangxi. The results of N gene sequencing and comparison showed that the 28 isolates and reference strains were classified into three groups, and most isolates belonged to group Ill, while the isolates in 1985-2008 belonged to groups IV and II. The data of VN test indicated that the 28 isolates belonged to 6 serotypes; among them, 71. 4% belonged to serotypes 1, 2, and 3, and 11 (39.3%) shared the same serotype with the current vaccine strains. Given the data of our previous study, it is found that prevalent serotypes and their proportions varied in different areas of Guangxi and during different periods. These data lay a good foundation for developing an oil-emulsified inactivated polyvalent vaccine containing local dominant serotypes for the effective prevention and control of infectious bronchitis.
Animals ; Antibodies, Viral ; immunology ; Chick Embryo ; Chickens ; China ; epidemiology ; Coronavirus Infections ; epidemiology ; immunology ; veterinary ; virology ; Infectious bronchitis virus ; classification ; genetics ; immunology ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; epidemiology ; immunology ; virology

OBJECTIVETo search new clues to reveal the action mechanism of inhaled anesthetics.

METHODSThree kinds of Drosophila melanogaster were used as studied models: the wild type strain (H), the sevoflurane-sensitive strain (S), and the sevoflurane-resistant strain (R). Differential display reverse transcriptional-polymerase chain reaction method was performed to examine the differentially expressed fragments between Drosophila induced with and without sevoflurane. Rapid amplification of cDNA ends (RACE) method was used to clone the full length cDNA from positive differentially expressed fragments.

RESULTSThirty-one differentially expressed fragments were found between the two groups. Three fragments were identified as the positive differentially expressed fragments by Northern blot analysis. Two full-length cDNAs were cloned by RACE method, among which one was a 1.0 kb Drosophila calmodulin (CaM), located on Chr.2; the other was a 4.1 kb gene with unknown function (No.45), located on Chr.3.

CONCLUSIONThe two full-length cDNAs belong to the genes that related to anesthetic action pathway, which might participate in the regulation of cellular functions and signal transduction pathways. The two genes that we found should provide a novel way to study the mechanism of inhaled anesthetic action.

Anesthetics, Inhalation ; pharmacology ; Animals ; Cloning, Molecular ; DNA, Complementary ; Drosophila melanogaster ; drug effects ; genetics ; Gene Expression ; Methyl Ethers ; pharmacology ; Signal Transduction

This study is to elucidate the metabolic pathway of 1,2-[bis (1,2-benzisoselenazolone-3 (2H)-ketone)]-ethane (BBSKE) in rats. Rats were administrated with a single dose of BBSKE 200 mg x kg(-1). The metabolites in rat urine, feces, bile and plasma were identified by LC-MSn analysis. The characterization of fragment ions from LC-MSn chromatography and mass spectrometry was applied to the investigation of structures of metabolites. Three phase I metabolites were detected in rat urine and feces. Two of them were also found in plasma and one existed in bile. These products were derived from oxidized, methylated and S-methylated BBSKE, separately. One phase II glucuronide of BBSKE was also found in bile. Therefore, it is possible that BBSKE was metabolized by oxidization, methylation and glucuronidation.
Administration, Oral ; Animals ; Antineoplastic Agents ; administration & dosage ; blood ; metabolism ; urine ; Bile ; metabolism ; Bridged Bicyclo Compounds, Heterocyclic ; administration & dosage ; blood ; metabolism ; urine ; Chromatography, Liquid ; Feces ; chemistry ; Male ; Organoselenium Compounds ; administration & dosage ; blood ; metabolism ; urine ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization