1.HIF-1α and malignant tumors of digestive system
Journal of International Oncology 2009;36(11):861-864
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.
2.Progress of ameliorating the neurotoxicity of anaesthetics on developing brain
Fudan University Journal of Medical Sciences 2017;44(4):532-537
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.
3.Development, validation and preliminary application of highperformance liquid chromatography coupled with charged aerosol detector method for determination of polysorbate 80 in romiplostim
Chinese Journal of Biologicals 2024;37(11):1361-1366
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.
4.Characterization analysis of capsid protein of recombinant adeno-associated virus 6 using reversed phase liquid chromatography-mass spectrometry
Chinese Journal of Biologicals 2024;37(07):769-774
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.
5.Evaluation of removal efficiency of human interferon α2b chromatography process for impurity proteins using liquid chromatography-high resolution mass spectrometry
Chinese Journal of Biologicals 2025;38(02):179-184
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.
6.The in vitro HAART pharmacodynamics study with dolutegravir as the "anchor".
Acta Pharmaceutica Sinica 2015;50(1):50-8
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.
7.Pure tone threshoIds and Otoacoustic Emissions in PCI Patients without Hearing Loss CompIaints
Journal of Audiology and Speech Pathology 2015;(1):34-36
Objective To study the audiometry and otoacoustic emissions in patients with posterior circulation ischemia(PCI) .Methods Forty patients treated by neurologists were selected as the experimental group who re_ceived pure tone audiometry ,tympanometry ,otoacoustic emissions (OAE) and distortion products otoacoustic emis_sions (DPOAE) examinations .Thirty healthy objects were chosen as the control group .The data from the PCI groups and the control group were compared using the SPSS 14 .0 software .ResuIts The difference in pure tone thresholds across 125 to 8 000 Hz between the PCI group and the control group was not statistically significant (P>0 .05) .In the PCI patients ,the prevalence of TEOAE was only 47 .5% ,significantly different from that of in the control group .The prevalence of DPOAE at 0 .5~8 kHz were between 57 .9% ~77 .6% in PCI patients ,and the amplitudes of DPOAE were reduced significantly (P<0 .05) .ConcIusion Cochlear damages can occur to patients with PCI ,especially in the high frequency range .These results suggest that OAE can be used as an important diag_nostic test for patients with PCI ,and might be helpful for the location diagnosis of PCI .
8.Sirolimus in kidney transplantation:theory and technology
Chinese Journal of Tissue Engineering Research 2014;(5):779-784
BACKGROUND:Calcineurin inhibitors reduce acute rejection rates and improve short-term graft survival in renal transplantation, but its nephrotoxicity associated with long-term use of calcineurin inhibitors remains an important issue. To both avoid exposure to calcineurin inhibitors and maintain effective immunosuppression, immunosuppressive agents such as sirolimus have emerged.
OBJECTIVE:To summarize the research progress of the two main protocols of sirolimus in kidney transplantation (de novo sirolimus-based therapy without calcineurin inhibitors and protocol conversion from a calcineurin inhibitor based therapy to sirolimus).
METHODS:With the key words of“kidney transplantation, sirolimus”in Chinese and in English, respectively, a computer-based search of articles was performed in CNKI (January 2000 to September 2013) and PubMed (January 1996 to September 2013) databases. Articles with the de novo sirolimus-based therapy without calcineurin inhibitors and protocol conversion from a calcineurin inhibitor based therapy to sirolimus were included.
RESULTS AND CONCLUSION:Sirolimus may obtain the advantages of no renal toxicity, anti-tumor and lower incidence of cytomegalovirus infections when compared with calcineurin inhibitors. But not al patients are suitable for sirolimus, and to screen patients strictly is the key of satisfactory clinical results. An appropriate treatment plan, drug monitoring of sirolimus, prevention and treatment of complications are essential features of the use of sirolimus.
9.EXPERIMENTAL STUDY ON THE EFFECT OF REMIFENTANIL ON THE EXPRESSION OF ADHENSIVE FACTORS IN LUNG INJURY IN RATS
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
To determine the effects of Remifentanil on the expression of adhesive factors during acute lung injury, we used immunohistochemistry and reverse transcription polymerase chain reaction (RT PCR) technics to measure the expression levels of ICAM 1, and P Selectin in the lung in a rat model of hemorrhagic shock. We duplicated modified Wigger hemorrhagic shock model in healthy SD rats. The rats were divided into three groups: control group, resuscitation group, resuscitation and drug using group. The positive ratios of ICAM 1 and P Selectin in the lungs at different time points, namely preshock, shock, resuscitation 2 hours, and resuscitation 4 hours were determined. There were 6 rats in each group and each time point. The degree of pulmonary injury in the drug using group was less than that in other two groups. Resuscitation with lactated Ringer's solution led to most serious injury in the resuscitation group. In drug using group, the expression of ICAM 1 and P Selectin was far less than that in the other two groups ( P
10.STUDY ON THE EFFECT OF REMIFENTANIL ON THE EXPRESSION OF APOPTOSIS FACTORS IN LUNG IN RATS WITH HEMORRHAGIC SHOCK
Medical Journal of Chinese People's Liberation Army 2001;0(10):-
Objective To determine the modulating effects of remifentanil on the expression of apoptosis factors during acute lung injury as a result of hemorrhagic shock, we observed the expression of C-Jun, Bax, Bcl-2 in lung with immunohistochemistry in rats with hemorrhagic shock. Methods Hemorrhagic shock model was replicated with modified Wigger's method in healthy SD rats. Seventy-two rats were randomized into three groups: sham resuscitation, resuscitation with lactated Ringer′s solution, and fluid resuscitation with remifentanil. Positive rates of C-Jun, Bax, Bcl-2 were assayed in the lungs at different time points: preshock, shock, 2 hours after resuscitation, and 4 hours after resuscitation. Results ①The model of acute lung injury subsequent to hemorrhagic shock was reproduced in rats. ②The expression of Bcl-2 elevated slowly in three groups, while the expression of Bax elevated rapidly, and there were significant differences between the value of baseline and those of other time points (P