General anesthetics are widely used in pregnant women,gravidas and infants.In the basic studies of rodents,mammals and non-human primate,general anesthetics can cause neurotoxicity,neuroapoptosis and damage neurodevelopment on the developing brain.Therefore,To explore protective measures and mechanism of anesthetic neurotoxicity is of great significance for formulating clinical anesthesia plan,guiding clinical obstetrics and pediatric anesthesia.This article reviewed the progress of ameliorating the neurotoxicity of general anaesthetics on developing brain including anesthetic assistants,hormone drugs,plant extracts,nutritional components and others.

Hie expression of hypoxia-inducible factor- 1a ( HIF-1a) is one of the important factors that cause most solid tumors rapid growth in hypoxic microenvironment, HIF-1a plays an important role in tumor angjogenesis, cell proliferation, metastasis and apoptosis. Similarly, HIF-1a also plays important biological roles in the occurrence and progression of digestive system carcinomas, such as esophagus cancer, stomach cancer, colon cancer, liver cancer and pancreatic cancer. HIF-1a can be used as one of the important molecular markers in prevention and treatment of gastrointestinal cancers.

Objective To characterize the capsid proteins of recombinant adeno-associated virus 6(rAAV6)vectors by reversed phase liquid chromatography-mass spectrometry(RPLC-MS),including primary structure and post-translational modification(PTM).Methods The mobile phase A consisted of 0. 1% aqueous solution of difluoroacetic acid(DFA),while the mobile phase B was 0. 1% DFA acetonitrile solution. The column temperature was maintained at 80 ℃,and the gradient elution lasted for 10 min(0→10 min,mobile phase B 15%→45%). The ESI-Q-TOF mass spectrometry detection operated in positive ion mode with the scanning range of 400-4 000 m/z,the scanning frequency of 2 Hz,the cone voltage at 80 V,the capillary voltage at 3. 0 kV,and the ion source temperature at 120 ℃.Results The measured relative molecular mass of the AAV capsid proteins VP1,VP2,and VP3 was 81 255. 9,66 062. 9,and 59 488. 6,respectively. The deviations from the theoretical values were 8. 1 ppm for VP1,3. 8 ppm for VP2,and 36 ppm for VP3. Mass peptide profile analysis of the enzymatically digested rAAV6 sample indicated a sequence coverage of about 89% with detected PTMs mainly including deamidation,N-terminal acetylation,ubiquitination,and phosphorylation;no glycosylation modification sites were found. Tandem mass spectrometry confirmed the N-terminal and C-terminal sequences of the rAAV6 capsid protein as well as the N-terminal PTM.Conclusion The complete relative molecular mass of rAAV6 capsid protein was analyzed by RPLCMS technique,and the PTM of rAAV6 capsid protein was analyzed by tandem mass spectrometry at the peptide level,which has a certain significance for the quality control of AAV gene therapy products and the improvement of production process.

Objective To develop, validate and preliminarily apply a high-performance liquid chromatography(HPLC)-charged aerosol detector(CAD) method for the quantitative analysis of polysorbate 80 in recombinant protein drug romiplostim, in order to provide a reference for the quality control of recombinant protein drug preparations.Methods The HPLCCAD determination method for polysorbate 80 content was developed by screening the mobile phase and optimizing the elution gradient and CAD parameters. The specificity, repeatability and accuracy of the method were validated, and the limit of detection(LOD), limit of quantitation(LOQ) and linear range were determined. The content of polysorbate 80 in two batches of romiplostim samples was determined by using the developed method.Results HPLC-CAD conditions after method optimization were as follows: Acclaim Surfactant Plus column(150 mm × 4. 6 mm, 3 μm) was adopted, and the column temperature was 25 ℃. The 0. 1% formic acid in water and 0. 1% formic acid in isopropanol were used as mobile phase A and B respectively, with a flow rate of 0. 6 mL/min and a gradient elution(0 min 80% A, 1. 8 min 80% A, 2. 0 min66. 5% A, 3. 0 min 66. 5% A, 3. 5 min 0% A, 8. 5 min 0% A, 9. 0 min 80% A, 15. 0 min 80% A). The injection volume was10 μL. The evaporative temperature for CAD was 50 ℃ and the power function value(PFV) was 1. 25. There was no interference of other chromatographic peaks in the sample matrix and blank solution in this method. The RSDs of peak area for repeatability at low(10 mg/L), medium(50 mg/L) and high(80 mg/L) concentrations were 4. 32%, 1. 97% and 3. 46%, and the mean spike recoveries were 95. 7%, 99. 0% and 98. 9%, respectively. Polysorbate 80 had a good linear relationship in the concentration range of 5. 0-100. 0 mg/L(R2= 1. 000 0), and the LOD and LOQ in the method were 1. 0 and 2. 0 mg/L respectively. The content of polysorbate 80 detected by the developed method was 33. 20 and 32. 28 mg/L in two batches of romiplostim samples respectively.Conclusion The developed HPLC-CAD method for the determination of polysorbate 80in romiplostim has high sensitivity, specificity, repeatability and accuracy, which can be used for the determination of polysorbate 80 in the quality control of recombinant protein drugs.

Objective To evaluate the removal efficiency of chromatography process for impurity proteins[IFN-related proteins and host cell proteins (HCPs)]in the purification process of human interferon α2b (h IFNα2b) by liquid chromatogra-phyhigh resolution mass spectrometry (LC-HRMS),in order to provide experimental basis for improving the production process of h IFNα2b.Methods The whole protein levels of IFN-related proteins (including deamidated IFN,oxidized IFN and acetylated IFN) and HCPs during the chromatography process of h IFNα2b were analyzed by LC-HRMS,and the h IFNα2b post-translational modification sites and HCPs were detected on the polypeptide levels.Results HCPs could be removed by hydrophobic interaction chromatography (HIC) M column,HCPs,deamidated IFN and acetylated IFN could be removed by anion exchange chromatography (AEC),and oxidized IFN could be removed by HIC (B column).The major acetylation sites (K34,K121,K131 and K134),major oxidation sites (M16,M21,M59 and M148),as well as Q5 and Q21 of major deamidation sites (N45,N156,Q5,Q21,Q46 and Q124) of h IFNα2b were located on the surface of the protein,which were susceptible to environmental conditions and subjected to acetylation and oxidation.Conclusion In the purification process of h IFNα2b,chromatography purification can effectively remove IFN-related proteins and HCPs,which also indicated that LC-HRMS technology can be used for the research of IFN purification process.

This study is to evaluate the HAART pharmacodynamics with dolutegravir as the "anchor" in vitro. A nucleoside reverse transcriptase inhibitors (NRTIs) resistant recombinant virus model (VSVG/HIV-1(RT-D67N,K70R,T215F)) and an integrase inhibitors (INIs) resistant recombinant virus model (VSVG/HIV-1(IN-G140S,QI48H)) were constructed and established. The anti-viral pharmacodynamics was evaluated with drug combinations including two NRTIs along with one INI or one NNRTI. The results showed that the combination with an INI gave a stronger synergism on wild type HIV-1 replication comparing to that with an NNRTI. Comparing the two INIs as the "anchor" for HAART, DTG exhibited an equivalent CI to that of RAL on wild type HIV-1 replication; but a greater synergy than RAL on INI-resistant HIV-1 replication. Besides of the pharmacodynamics results of DTG-based drug combination, the results may contribute to clinical antiviral therapy.

Adult ; Biomarkers, Tumor ; metabolism ; Calbindin 2 ; Female ; Follow-Up Studies ; Humans ; Mesothelioma ; metabolism ; pathology ; surgery ; Mucin-1 ; metabolism ; Peritoneal Neoplasms ; metabolism ; pathology ; surgery ; S100 Calcium Binding Protein G ; metabolism

This study is to evaluate the HAART pharmacodynamics with dolutegravir as the "anchor" in vitro. A nucleoside reverse transcriptase inhibitors (NRTIs) resistant recombinant virus model (VSVG/HIV-1(RT-D67N,K70R,T215F)) and an integrase inhibitors (INIs) resistant recombinant virus model (VSVG/HIV-1(IN-G140S,QI48H)) were constructed and established. The anti-viral pharmacodynamics was evaluated with drug combinations including two NRTIs along with one INI or one NNRTI. The results showed that the combination with an INI gave a stronger synergism on wild type HIV-1 replication comparing to that with an NNRTI. Comparing the two INIs as the "anchor" for HAART, DTG exhibited an equivalent CI to that of RAL on wild type HIV-1 replication; but a greater synergy than RAL on INI-resistant HIV-1 replication. Besides of the pharmacodynamics results of DTG-based drug combination, the results may contribute to clinical antiviral therapy.
Antiretroviral Therapy, Highly Active ; Cells, Cultured ; Drug Resistance, Viral ; HIV Integrase Inhibitors ; pharmacology ; HIV-1 ; drug effects ; physiology ; Heterocyclic Compounds, 3-Ring ; pharmacology ; Humans ; Virus Replication ; drug effects

Although small molecule drugs (SMD) are still mainstream for the treatment of diseases, large molecule biologicss of many advantages, pose a challenge to the further discovery and use of SMD. The advantages of SMD are the convenience of oral administration and good patient compliance. However, the challenge with SMD is to integrate the PD, PK, selectivity and safety into a chemical structure. Because of their small size and surface area they often bind to various proteins, and off-target actions can cause adverse reactions. In this sense, selectivity is critical. Based upon target as the core to construct a chemical structure, it is necessary to consider the requirements of all the attributes, but achievement of the full-dimensional optimization is difficult. Modern drug discovery has been greatly enhanced by molecular biology and structural biology, and new strategies and technologies have emerged, which have created many successful medicines. For example, under the guidance of structural biology, covalent binding drugs connect moderate "electrophilic warheads" to the appropriate positions of molecules, and upon binding to their targets the electrophiles are irreversibly linked to the target by covalent bonds. Molecular biology can be directly applied to the development of antibody-coupled drugs (ADC). The antibody (A) acts as a carrier and a guide (for PK), and carries toxic molecules (D) into cancer cells, thus playing a killing role (for PD). The separate pharmacodynamic and pharmacokinetic entities are coupled (C) by linkers. PROTACs are also bifunctional molecules, which recruit a target protein and ubiquitin ligase E3 to form a ternary complex, which then acts as a catalyst to ubiquitinate the target protein and lead to degradation by the proteasome. In addition, in recent years, the combination of two fixed-dose drugs has improved selectivity, safety, and long-term benefit with many severe diseases, and can be regarded as an innovative strategy of physical combination. This review discusses some successful examples to briefly present the principles from the perspective of medicinal chemistry and therapeutic application.

Antibodies, Viral ; analysis ; Cerebrospinal Fluid ; virology ; Child ; Child, Preschool ; Feces ; virology ; Genotype ; Humans ; Infant ; Meningoencephalitis ; diagnosis ; virology ; Parechovirus ; classification ; genetics ; isolation & purification ; Picornaviridae Infections ; diagnosis ; epidemiology ; virology ; RNA, Viral ; genetics ; Respiratory Tract Infections ; diagnosis ; virology ; Sepsis ; diagnosis ; virology ; Sequence Analysis, DNA