Establishment of a genetic monitoring method for laboratory quails / 中国比较医学杂志

Objective To establish a genetic monitoring method for laboratory quails.Methods Quail microsatellite loci were searched in the literature,and microsatellite DNA loci suitable for quails were screened by an interspecific transfer method in closely related species,namely chickens and ducks.Quail liver DNA was extracted as a template,and the corresponding loci were screened by PCR amplification and agarose gel electrophoresis.On the basis of amplification of the selected microsatellite loci,the number of alleles,polymorphisms,and microsatellite loci combinations for quail genetic quality detection were selected and detection method were developed.Results We preliminary determined 23 microsatellite loci for genetic monitoring of closed-colony laboratory quails.Conclusions A genetic monitoring method for laboratory quails was preliminary developed.

Study on the Antibody Production Efficiency in Modified Big-BALB/c Mice / 实验动物与比较医学

Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies inducedin vivo through hybridoma.Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.ResultsAfter 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×104 U and 33.22×104 U) was higher than that of BALB/c mice (5.33×104 U and 19.31×104 U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×104 U/g) was higher than that of BALB/c mice (0.23×104 U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (bothP＜0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation.

Excavation and evaluation of adverse reaction signals of 4 kinds of imported PD- 1/PD-L1 inhibitors / 中国药房

OBJECTIVE To exc avate and evaluate the adverse reaction signals of 4 kinds of imported programmed cell death 1 （PD-1）and its ligand （PD-L1）inhibitors，and to guide rational drug use in clinic. METHODS OpenVigil 2.1 software was used to obtain the adverse event reports of four drugs as nivolumab ，pembrolizumab，atezolizumab and durvalumab from the first quarter of 2013 to the fourth quarter of 2020 in the US FDA adverse event reporting system. The reporting odds ratio and proportional reporting ratio were used for signal mining to evaluate new or potential adverse reaction signals. RESULTS A total of 46 840 reports of adverse events with PD- 1/PD-L1 inhibitors as the primary suspected drug were collected ，including 28 896 reports of nivolumab，13 298 reports of pembrolizumab ，3 398 reports of atezolizumab ，and 1 248 reports of durvalumab. From the general characteristics of these reports ，the gender distribution was more men than women ，and the age distribution was mainly in the range of 51-85 years old. The reporting year was mainly in the nearly 4-5 years，and the main reporting countries were the US and Japan，with“death”and“hospitalization or prolonged hospitalization ”as the main serious adverse events which were over 50％ of the whole of 4 kinds of adverse events. A total of 1 597 adverse reaction signals were obtained ，involving 26 systems，focusing on “benign，malignant and unspecified neoplasms （cystic and polypoid tumor ）”，“infections and infestations ”and“investigations”， etc. The analysis of the top 50 adverse reaction signals showed that the largest number of report was endocrine system disease ，the most frequency signal was “malignant neoplasm progression ”and the strongest adverse reaction signal was “radiation pneumonitis ”. And it was also found that 13 adverse reaction signals ，such as “radiation pneumonitis ”“cholangitis”“fulminant type 1 diabetes mellitus” “blood creatine phosphokinase increased ” “disseminated intravascular coagulation ”“cardiac failure ”and “cerebral infarction ”，etc.，were not reported in the drug instructions. CONCLUSIONS PD-1/PD-L1 inhibitors mediate a large number of adverse reaction signals，resulting in high safety risks in “benign，malignant and unspecified neoplasms （cystic and polypoid tumor ）”，“infections and infestations ”and “investigations”，etc. The newly discovered 13 adverse reaction signals ，such as “radiation pneumonitis ”“cholangitis”“blood creatine phosphokinase increased ”“cardiac failure ”and“cerebral infarction ”are of great significance for guiding rational drug use in clinic.

Establishment of a detection method for Helicobacter pylori in living Mongolian gerbil / 中国比较医学杂志

Objective To establish a detection technique for H.pylori(HP) infection in Mongolian gerbils using nested PCR technique.Methods H.pylori was cultured in vitro and inoculated into Mongolian gerbils.At the 10th week after infection, the HP in the gastric juice of Mongolian gerbil was detected by conventional PCR assay and the gastric juice, gastric mucosa, duodenal contents and colon stool were examined by nested PCR.Rapid urease test and ELISA were used to analyze the accuracy of the nested PCR assay.All of the PCR products were verified by sequencing.Results The positive rate of gastric juice detected by conventional PCR was 30%, while the positive rates of gastric juice, gastric mucosa, duodenal contents and colon stool detected by nested PCR were 100%, 100%, 90%, and 10%, respectively.The positive detection rates of rapid urease test and serum ELISA were 100% and 0%, respectively.Comparing the results of different methods, both the positive rates of gastric juice and gastric mucosa detected by nested PCR and the detection rate of rapid urease test were 100%, but the results of conventional PCR detection of gastric juice, the nested PCR detection result of stool in colon and of serum ELISA assay were lower than other methods.Conclusions Due to its high accuracy and sensitivity, the nested PCR assay of gastric juice can be used for the long-time detection of H.pylori infection in Mongolian gerbils, especially useful in the experiments of prevention and treatment of H.pylori infection.

Genetic quality analysis of 24 domestic inbred mouse strains by microsatellite DNA / 中国比较医学杂志

Objective To analyze the genetic quality of 24 domestic inbred strains mice using microsatellite loci panel.Methods Previously selected 30 microsatellite loci of mouse with high polymorphism and more allele numbers were used to synthesize corresponding fluorescently-labeled primers.Then the genomic DNA samples of each mouse were amplified by PCR and the products were analyzed by STR scanning to genotype the inbred strains of mice.Results Out of the 24 inbred strains, 15 inbred strains showed the same genotype within one strain at 30 loci.Among different strains, microsatellite loci indicated polymorphism which could be used to distinguish different strains.However, the rest 9 strains demonstrated polymorphism within strains.Conclusions Our stuoly provides a useful microsatellite panel to detect genetic quality of inbred mice and distinguish different strains with the optimized microsatellite loci.

The optimization scheme of Mongolian gerbils superovulation induced by PMSG and hCG / 中国比较医学杂志

Objective To optimize the scheme of Mongolian gerbil superovulation.Methods Based on the analysis of the animal-week-old, the dose and time interval of hormone, we got the best animal-week-old, dose and time interval of hormone for Mongolian gerbil superovulation.Results The 6 week old female Mongolian gerbils which injected of 10 IU PMSG and followed by 10 IU hCG in 70 hours later could get the best superovulation.We collected eggs at 16 hour after mate with male gerbils.The ovum pick-up rate reached 80%, the number of oocytes were 32.6 ±3.0, the number of the fertilized egg developed to 2-cell are 24.8 ±5.4.Conclusions This study summarizes the optimization scheme of Mongolian gerbil superovulation induced by PMSG and hCG and it supported the foundation for Mongolian gerbils embryo biotechnology.

Advances in development and application of Mongolian gerbil resource / 中国实验动物学报

Mongolian gerbil is a multifunctional laboratory anima1 resource with Chinese characteristics.It has many advantages for some research fields, and play an important role.The more biological characteristics of Mongolian ger-bils discovered with in-depth research will promote diverse application of this resoure.In this paper, we reviewed the devel-opment of application of Mongolian gerbils in taxonomy, parasitology, microbiology, cerebral ischemia, lipid metabolism, neurological diseases, and cancer researches.

Transgene complete silencing may associate with rearrangement of retroviral vector / 生物医学工程学杂志

Transgene silencing is one of two major obstacles in both basic biomedical research for transgene and clinical practice of gene therapy. Based on the model of HT1080 cell clones, which transduced single copy of retroviral vector MGPN2, the mechanism of transgene silencing was explored in this investigation by a serial molecular techniques. In the HT1080 cell clone that absence of GFP protein synthesized, no significant aberration of epigenetic modification was detected, but the transcript size and the sequence changed that resulted in the reading frame shift. In addition to chromosomal position effect leading to transgene silencing, the transcript reading frame shift associated with retroviral vector rearrangements could induce complete silencing of transgene.

Comparison of domestic idataubicin and imported daunorubicin on the treatment of acute leukemia / 白血病·淋巴瘤

Objective To compare the effect and toxicity of domestic idarubicin (IDA) and imported daunorubicin (DNR) in the treatment of acute leukemia (AL).Methods According to the random number table method,68 patients were randomly divided in IDA group with 35 patients and DNR group with 33 patients.In IDA group,the patients with acute myelocytic leukemia were treated following IA scheme (domestic idataubicin plus cytosine arabinoside) and the patients with acute lymphoblastic leukemia were treated following VICLP scheme (vincristine,domestic idataubicin,cyclophosphamide,lasparaginase and prednisone).In DNR group,the patients with acute myelocytic leukemia were treated following DA scheme (imported daunorubicin plus cytosine arabinoside) and the patients with acute lymphoblastic leukemia were treated following VDCLP scheme (vincristine,imported daunorubicin,cyclophosphamide,lasparaginase and prednisone).Results In IDA group,21 patients achieved a complete remission(CR),5 patients achieved a partial remission(PR),with a 74.2 ％ (26/35) remission rate (RR).In DNR group,the remission rate was 62.3 ％ (20/33).No differences of the remission rate was found between the two groups (t =0.89,P =0.50).17 patients were found remission over one year in IDA group,and 6 patients were in DNR group.The difference was statistically significant between the two groups (x2 =5.56,P =0.02).Conclusion IDA is more effective than DNR in AL treatment.The higher RR and longer remission time are found in IDA group than DNR group.IDA is effective and safe in the treatment of AL.