Objective To establish the drug resistant cell line of choriocarcinoma and to study the transfection of the human interleukin 2 (hIL 2) gene into the established drug resistant cell line and investigate the reversal of the multidrug resistance Methods The resistant cell line was established by pulse exposed choriocarcinoma cell line JEG 3 to etopside (VP 16) for ten months The recombinant plasmid containing pcDNA3 1(+) hIL 2 gene was constructed The drug resistant cell line was transfected with the constructed plasmid by lipofectin, and the tumor cell colonies containing the IL 2 sequence were selected by genetin The expression of hIL 2 and drug resistant related genes was detected by reverse transcript polymerase chain reaction The chemosensitivity of the gene transfected tumor cells and the non transfected cell lines to methetraxate, VP 16, kengshengmycine, paclitaxol and 5 fluorouracil was determined by the methyl thiazolyl tetrazolium cytotoxicity assay Results The transfected cells expressed human hIL 2 gene, and showed the reversal of multidrug resistance by methyl thiazolyl tetrazolium assay The transfected cells expressed no multidrug resistance gene 1 (MDR1) on mRNA level Drug resistance index to VP 16 decreased from 38 7 to 6 0 and 6 1, the index to methetraxate decreased from 14 5 to 2 6 and 2 5, to methetraxate from 13 0 to 2 0 Conclusion The transfection of hIL 2 gene into the drug resistance cell line of choriocarcinoma can modulate the MDR1 expression on the mRNA level, and reverse the drug resistance

AIM: To explore the feasibility of culturing human umbilical vein endothelial cells ( HUVEC ) on acellular corneal stroma and performing the posterior lamellar endothelial keratoplasty ( PLEK ) treating corneal endothelial decompensation.
METHODS: Thirty New-Zealand rabbits were divided into three groups randomly, 10 rabbits for experimental group, 10 for stroma group and 10 for control group. Corneal endothelial cells were removed to establish animal model of corneal endothelial failure. PLEK was performed on the rabbits of experimental group and stroma group, and nothing was transplantated onto the rabbits of control group with the deep layer excised only. Postoperative observation was taken for 3mo. The degree of corneal edema and central corneal thickness were recorded for statistical analysis.
RESULTS:Corneas in experimental group were relieved in edema obviously compared with that in stroma group and the control group, and showed increased transparency 7d after the operation. The average density of endothelial cells was 2 026. 4±129. 3cells/mm2 , and average central corneal thickness was 505. 2±25. 4μm in experimental group, while 1 535. 6±114. 5μm in stroma group and 1 493. 5±70. 2μm in control group 3mo after operation.
CONCLUSION:We achieved preliminary success in our study that culturing HUVEC on acellular corneal stroma and performing PLEK for corneal endothelial decompensation. HUVEC transplanted could survive in vivo, and have normal biological function of keeping cornea transparent. This study provides a new idea and a new way clinically for the treatment of corneal endothelial diseases.

Tumorigenesis is a complex process that is regulated by a variety of network signals. With the continuous development of the process, tumor cells gradually exhibit lots of hallmarks.Tumor cells have the characteristics of unlimited proliferation, resistance to apoptosis, evading immune surveillance, among others. As a unique organelles, mitochondria play an important role in cellular energy metabolism, reactive oxygen species producing and apoptosis process. Particularly, mitochondria have a close relationship with tumor development. In this review, we focus on the essential role of mitochondria in tumor cells development.
Animals ; Energy Metabolism ; Humans ; Mitochondria ; metabolism ; physiology ; Neoplasms ; etiology ; genetics ; physiopathology ; Tumor Microenvironment ; physiology

Objective To observe the electrocardiogram (ECG) of urban residents in non-endemic areas of Keshan disease in Ji'nan City,Shandong Province,and to provide scientific basis for health evaluation and prevention and control of Keshan disease.Methods From July 2014 to June 2015,ECGs of residents undertaken healthy examination in Shandong Ciming Physical Examination Center were collected,retrospective analysis was used to analyze residents' heart health.Statistical analysis was conducted according to sex and age groups,and the diagnosis of ECG was based on Concise Pediatric Electrocardiogram and Clinical Electrocardiography.Results Totally 31 753 ECGs were recorded,including 18 664 males accounting for 58.8％ and 13 089 females accounting for 41.2％,who were aged 6-78 years old,including 304 people 6-20 years old,18 861 people 21-40 years old,10 451 people 41-60 years old,and 2 137 people 61-78 years old.Totally 27 699 normal ECG (87.2％) and 4 054 abnormal ECG (12.8％) were detected,there were 2 212 males and 1 842 females in the abnormal ECG,the detection rates were 11.9％ (2 212/18 664) and 14.1％ (1 842/13 089) for males and females,respectively.The detection rate of female was significantly higher than that of male (x2 =34.082,P ＜ 0.01).The detection rate of ST-T change in abnormal ECG was the highest,which was 316.5/10 000 (1 005/31 753),men and women were 273.3/10 000 (510/18 664) and 378.2/10 000 (495/13 089),respectively.The detection rate of arrhythmia was 4.8％ (1 523/31 753),including 1 023 males and 500 females,and their detection rate of arrhythmia was 548.1/10 000 (1 023/18 664) and 382.0/10 000 (500/13 089),respectively.In arrhythmia,degree Ⅰ atrioventricular block (155.3/10 000,493/31 753),complete rightbundle branch block (81.6/10 000,259/31 753),ventricular premature beats (79.0/10 000,251/31 753),and atrial premature beats (48.2/10 000,153/31 753) had higher detection rates.Conclusion The detection rate of women of urban residents in Ji'nan is higher than that of men,the most common abnormal ECG is ST-T change,and the detection rate of arrhythmia is low.

Objective To study the healing response of a tissue engineered bone of bMSCs combined rFN/CDH-BCP in a rabbit model. Methods The biomimetic surface was achieved by immobilizing rFN/CDH onto biphasic calcium phosphate ceramic ( BCP) covalently. The effect of rFN/CDH-BCP on adhesion,proliferation and differentiation were evaluated comprehensively by using cell centrifugal adhesive as-say,MTT method,SEM,Alkaline phosphatase ( ALP) activity and alizarin red staining respectively. A rabbit lumbar fusion model was estab-lished by implanting bMSCs combined rFN/CDH-BCP into intertransverse process space of L4 ~L5 ,the fusion site bone formation was ob-served by imaging means,histological techniques was used to observe the new bone formation and distribution of seed cells. Results Cell centrifugal adhesive assay indicated the adherent bMSCs on rFN/CDH-BCP associated with the ligand density,the biomimetic surface posses-ses excellent biocompatibility. The ALP activity on rFN/CDH-BCP surface was the highest among all samples on the 10th day after induction (P<0. 05). On the 21st day,alizarin red staining showed that the oval-shaped and orange-red nodules,either the number or the area,distrib-uted wider on rFN/CDH-BCP surface. The results from X plain after 3 months revealed a fuzzy gap between material and bone bed,and higher cover rate of intertransverse process space with new bone deposition on rFN/CDH-BCP surface. Conclusion Histologically,rFN/CDH-BCP exhibited as interlacing bone trabecula bridging biomaterial suface and cortical bone of transverse process continously. The bone mass was much more than the ture BCP class. The comprehensive data reveals that when loaded with MSCs,rFN/CDH-BCP demonstrates superior char-acteristics of osteoconduction and osteoinduction,and substantially enhances healing capacity in vivo.

Objective To study the effect of Danggui Buxue Tang (DBT) on immunological reconstitution of bone marrow transplantation (BMT) in mice. Methods BALB/c mice were irradiated by 137Cs γ once 8.5 Gy, then the mice were engrafted with bone marrow cells (107 cells/mouse) within 4 hours lethal irradiation. And the mice were fed by DBT every day for 15 days. Flow cytometry technique combined with immunological methods were performed to evaluate immunological reconstitution of BMT mice in 30 and 60 days pest-transplantation. Peripheral blood RBC and WBC were counted, and nucleated cells were assayed in recipient bone marrow. Lymphocyte numbers in thymus and periphery were counted and subpopulations of the two origins were observed respectively. Lymphocyte proliferation induced by Con A and LPS, plaque-forming cell (PFC), delayed type hypersensitivity (DTH) were also examined in 30,60 days in post-transplantation respectively. Results Compared with BMT mice, BMT mice treated with a certain dose of DBT could increase the number of peripheral blood RBC and WBC in the recipients, and also could increase that of nucleated cells significantly. BMT mice treated with DBT could increase lymphocyte numbers in thymus and periphery, and improve thymocyte subpopulations, resulting in enhancement in immune function. Conclusion DBT can enhance the immunological reconstitution of BMT mice.

Objective To systematically review the existing evidence about the effect of medial and lateral (crossed) entry pins versus only lateral entry pin fixation on the supracondylar fractures of the humerus in children.Methods Eligible studies were identified in Cochrane library,the Cochrane Bone,Joint and Muscle Trauma Group (till March 2011 ),Medline (from 1966 to March 2011 ),EMbase (from 1966 to 2011 ),CBM (from January 1979 to March 2011 ),PubMed,Wanfang Data (from 2000 to March 2011 ) and CNKI and the references of the included studies and several Chinese orthopedic journals were manually searched.Then,the randomized controlled trials (RCTs) and quasi-randomized controlled trials (CCTs) about two entries (crossed and only lateral pinning) for supracondylar fractures of humerus in children were collected.After evaluation of methodology with the enrolled studies,available data was extracted and systematic review was conducted via the method recommended by the Cochrane Collaboration.Results In total,five RCTs involving 311 patients were involved.Compared with the preoperative data,the meta-analysis results showed no significant difference in reduction stability in terms of change in Baumann angle and Carrying angle between the two groups.For the postoperative function outcome including complete reduction,Flynn grade and full return to function,no significant difference was found between the two groups.For the postoperative complications,there was no difference in the infection of pin tract,though lateral entry resulted in a significant lower incidence of the iatrogenic nerve injury compared with the medial and lateral entry.Conclusions With the Kirschner wire fixation for supracondylar fractures of humerus in children,current existing evidences indicate that the lateral entry of pinning has similar results in reduction stability,function outcome and incidence of pin tract infection compared with medial and lateral entry.Nevertheless,lateral entry,as a safe pinning technique,may effectively decrease the risk of iatrogenic nerve injury

Objective To establish the m ethod to analyze γ-hydroxybutyric acid (GHB ) and its precursors 1,4-butanediol (1,4-B D ) and gam m a-butyrolactone (GBL) in urine through LC-M S/M S and provide evi-dence for related cases. Methods GHB-d6 and M O R-d3 were used as the internal standard. The urine sam ple w as separated by LC after protein precipitation w ith m ethanol. The electrospray ion source w as for ionization. E ach com pound w as detected through m ultiple-reaction m onitoring (M R M ) m ode. Results The lim its of detection of GHB and its precursors 1,4-B D and GBLwere 0.1, 0.1 and 2μg/m L. The accuracy w as 87.6% -98.1% . The intra-day and inter-day precisions were less than 15% and m atrix ef-fects were higher than 80% . Conclusion The m ethod is high sensitive, sim ple, rapid, specific and w ith high reliability. This study has provided technical support and basic data for forensic cases involving GHB .

Objective:To investigate the clinical and neuroimaging features of atypical teratoid/rhabdoid tumors (AT/RT) of the central nervous system and the survival prognosis in different locations.Methods:The clinical data of 27 patients with AT/RT confirmed by biopsy or postoperative pathology in Sanbo Brain Hospital of Capital Medical University from October 2012 to September 2020 were collected, including 17 males and 10 females, aged (7.6±10.0) years, ranging from 0.2 to 39.0 years old.The clinical features and the results of the first preoperative imaging examination were retrospectively analyzed.The patients were divided into supratentorial, infratentorial and trans supratentorial and infratentorial groups according to the location.The survival time of the three groups was compared by Kaplan-Meier survival curve.Results:All patients presented with headache, including 12 cases with vomiting.There were 12 cases of supratentorial, 10 cases of infratentorial and 5 cases of supratentorial and infratentorial.There were 20 cases of cystic degeneration, 7 cases of calcification, 6 cases of hemorrhage and 13 cases of peritumoral edema.The median survival time of patients with infratentorial AT/RT was longer than that of patients with supratentorial and transtentorial AT/RT (χ 2=7.353, P=0.025). Conclusion:Central nervous system AT/RT is easy to occur in young children, and the survival time of AT/RT patients is longer.

OBJECTIVE: To establish the method to analyze γ-hydroxybutyric acid (GHB) and its precursors 1,4-butanediol (1,4-BD) and gamma-butyrolactone (GBL) in urine through LC-MS/MS and provide evidence for related cases. METHODS: GHB-d6 and MOR-d3 were used as the internal standard. The urine sample was separated by LC after protein precipitation with methanol. The electrospray ion source was for ionization. Each compound was detected through multiple-reaction monitoring (MRM) mode. RESULTS: The limits of detection of GHB and its precursors 1,4-BD and GBL were 0.1, 0.1 and 2 μg/mL. The accuracy was 87.6%-98.1%. The intra-day and inter-day precisions were less than 15% and matrix effects were higher than 80%. CONCLUSION The method is high sensitive, simple, rapid, specific and with high reliability. This study has provided technical support and basic data for forensic cases involving GHB.
4-Butyrolactone/urine* ; Butylene Glycols/urine* ; Chromatography, Liquid ; Forensic Sciences ; Humans ; Hydroxybutyrates/urine* ; Mass Spectrometry ; Reproducibility of Results ; Tandem Mass Spectrometry