Objective To study the effect of cumulus cell apoptosis on the structure of human oocyte cultured in vitro and to probe into the reason why cumulus cell apoptosis rate can be used clinically to predict the developmental potential of oocyte. Methods Cumulus-enclosed oocyte complexes (COCs) at the GV stage were cultured in vitro. The apoptosis rate of cumulus cells for every COC was evaluated with HE staining, DAPI staining and TUNEL labeling methods. Oocytes were divided into two groups according to their cumulus cell apoptosis rate. Structure of oocytes was observed under light microscope and transmission electron microscope. Mature oocytes at MⅡ stage were fertilized and cultured in vitro for two days. Then the embryos were evaluated according to their morphology under light microscope.Results The oocytes with low cumulus cell apoptosis rate had a normal structure and a promising developmental potential, while the oocytes with high cumulus cell apoptosis rate had deformed structures and a poor developmental potential. Some deformed oocytes had uneven perivitelline space. For oocytes with a small perivitelline space, the organelle development was slow compared with those with a large perivitelline space. Some had unevenly piled organelles. Secondary lysosomes and large space probably caused by the decopmounding of lysosomes were found in the ooplasm of these oocytes. The deformed oocytes appeared to have lots of swollen mitochondria with blurred cristae and membrane. Some of these oocytes had fractured first polar bodies and abnormally thick or thin zona pellucida. The cell junctions and microvilli of cumulus cells reduced as well. Conclusion The effect of cumulus cell apoptosis on the structure of oocyte cultured in vitro was revealed. The reason why cumulus cell apoptosis rate prognosticated the developmental potential of oocyte was demonstrated in the aspect of oocyte structure.

To express a gene of heat shock protein 70(HSP70) in E.coli , HSP70 gene was amplified by PCR. The PCR products were cloned into pUCm T vector and were sequenced; The HSP70 gene was subcloned into vector pET 21a(+) and expressed in E.coli. SDS PAGE and Western blot were employed to identify the expression of the HSP70 gene. Results showed that a fragment about 1 96kb was amplified by PCR. Sequence analysis revealed that the sequence of HSP70 was correct. The HSP70 gene was cloned into pET 21a(+) identified by enzyme digestion and PCR. SDS PAGE and Western blot showed that a M 72 000 protein was expressed and could be recognized by anti HSP70 antibody. Therefore,HSP70 gene has been successfully expressed in E.coli .

Objective To construct a fusion gene vaccine of gastric cancer MG7-Ag mimotope and heat shock protein 70, and to investigate the humoral and cellular immune response in mice induced by the vaccine. Methods The coding sequence of MG7 mimotope was incorporated with HSP70 gene at the 3' terminus by PCR amplification. Then the PCR product was cloned into pcDNA3.1(+) vector to construct the eukaryotic expression vector. The pcDNA3.1/MG7+hsp70 was sequenced to ensure the proper encoding. C57BL/6 mice were immunized with the fusion gene vaccine, and pcDNA3.1/hsp70 and empty pcDNA3.1 were used as controls. The serum was collected at the 3rd week after each immunization. Cellular ELISA was performed to evaluate the induced humoral immunity. The splenocytes were first separated at the 9th week for the assay of antigen specific CTL lysis activity by 51Cr release. Results A fragment about 2.0 kb was obtained by PCR amplification. Sequence analysis revealed that the sequence of mimotope was connected successfully to 3' terminus of hsp70 and the fusion gene was cloned into the pcDNA3.1(+) successfully. Cellular ELISA results suggested that the serum level of MG7-Ag antibody appeared in the vaccinated mice at 9th week, while no MG7-Ag antibody was detected in the controls. The results of the specific lysis rate of splenocytes showed no statistical difference between fusion gene vaccine group and control groups. Conclusion The fusion gene vaccine was constructed successfully and the specific humoral immune response was induced by the fusion gene vaccine.

Objective To study the relationship between gastric bile reflux and the digestive symptoms. Methods The extent of bile reflux into the stomach during 24 hours was monitored by using ambulatory bilirubin monitoring technique in 76 persons among them 44 were complaining of digestive symptoms and 22 asymptomatic. Results The total time percentile of bile reflux,the total area of bile reflux and frequency of bile reflux in asymptomatic group and symptomatic group were 15 6%?14 0%, 20 6?18 7, 22 3 times?13 6 times, and 26 4%?21 3%, 48 7?60 8, 42 9 times?44 5 times. Conclusion Bile reflux into the stomach was significantly more frequent and intensive in symptomatic group compared with asymptomatic group

The treatment of chronic myeloid leukemia (CML) was revolutionized with the advent of the first-generation tyrosine kinase inhibitors (TKIs), but drug resistance developed during treatment, leading to the development of the second-generation (dasatinib, nilotinib, and bosutinib) and third-generation (ponatinib) TKI. Compared with previous treatment regimens, specific TKI can significantly improve the response rate, overall survival rate and prognosis of CML. Only a few patients with BCR-ABL mutation are insensitive to the second-generation TKIs, so it is suggested to select the second-generation TKIs for patients with specific mutations. For patients with other mutations and without mutations, the second-generation TKI should be selected according to the patient's medical history, while the third-generation TKIs should be selected for mutations that are insensitive to the second-generation TKIs, such as T315I mutation that is sensitive to ponatinib. Due to different BCR-ABL mutations in patients with different sensitivity to the second and third-generation TKIs, this paper will review the latest research progress of the efficacy of the second and third-generation TKIs in CML patients with BCR-ABL mutations.
Humans ; Antineoplastic Agents/pharmacology* ; Dasatinib/pharmacology* ; Drug Resistance, Neoplasm/genetics* ; Fusion Proteins, bcr-abl/genetics* ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy* ; Mutation ; Protein Kinase Inhibitors/therapeutic use*

OBJECTIVETo explore the clinical effects of surgical treatment with cable dragged reduction and cantilever beam internal fixation by posterior approach for odontoid fracture associated with atlantoaxial dislocation.

METHODSThe clinical data of 12 patients with odontoid fracture associated with atlantoaxial dislocation from January 2008 to December 2013 were retrospectively analyzed. There were 8 males and 4 females, ranging in age from 21 to 53 years with an average of 37.2 years. Eleven cases were fresh fracture and 1 case was old fracture, all patients complicated with atlantoaxial anterior dislocation. According to Anderson-D' Alonzo typing method modified by Grauer, 3 cases were type IIA, 5 cases were type IIB, 3 cases were type IIC, and 1 case was type IIIA. All patients underwent surgical treatment with cable dragged reduction and cantilever beam internal fixation by posterior approach. JOA score and ADI method were respectively used to evaluate the nerve function and reductive condition of atlantoaxial dislocation.

RESULTSAll patients were followed up from 6 months to 2 years with an average of 1 year and 3 months. At 1 week, 6 months after operation, and final follow up, JOA scores were 13.2±1.3, 13.5±1.4, 14.3±1.5, respectively, and these data were obviously better than that of preoperative 8.3±1.4(<0.05). Postoperative X rays and CT showed satisfactory reduction of atlantoaxial dislocation. At 1 week, 6 months after operation, and final follow up, ADI were (2.2±0.4), (2.4±0.6), (2.3±0.5) mm, respectively, and these data were obviously better than that of preoperative.(5.8±1.2) mm(<0.05). All screws and cables had good location without looseness and breakage, and bone graft got fusion.

CONCLUSIONSSurgical treatment with cable dragged reduction and cantilever beam internal fixation by posterior approach for odontoid fracture associated with atlantoaxial dislocation is a good method, with advantage of firm fixation and high safety. It could obtain good clinical effects.