To clone the murine alpha-fetoprotein (AFP) gene, construct the eukaryotic expression vector of AFP and express in CHO cells, total RNA were extracted from Hepa 1-6 cells, and then the murine alpha-fetoprotein gene was amplified by RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant of vector was identified by restriction enzyme analysis and sequencing. After transient transfection of CHO cells with the vector, Western blotting was used to detect the expression of AFP. It is concluded that the 1.8 kb murine alpha-fetoprotein gene was successfully cloned and its eukaryotic expression vector was successfully constructed.
CHO Cells ; Cloning, Molecular ; Cricetinae ; DNA, Complementary ; Eukaryotic Cells/metabolism ; Genetic Vectors ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; alpha-Fetoproteins/*biosynthesis ; alpha-Fetoproteins/genetics

Objective: To explore the monomer compounds in the seeds of Momrodica cochinchinensis and to study the differentiation of mouse melanoma B16 cells induced by p-hydroxylcinnamaldehyde (PHC). Methods: After being treated by five kinds of compounds [PHC, coniferylaldehyde, p-hydroxylbenzaldehyde (PHB), 3-O-methoxyaniline-p-hydroxylbenzaldehyde, and ligballinol] for 48 h, the inhibitory rate of B16 cell growth was measured by sulforhadamine B (SRB); Morphological changes of B16 cells induced by PHC for 24, 48, and 72 h were observed by Giemsa staining and phase contrast microscope; Melanin content and the activity of tyrosinase in B16 cells 48 h after the administration were assessed by colorimeter. The expression of tyrosinase mRNA was detected by RT-PCR. Results: All the five compounds had the inhibitory effect on the B16 cells. Among them, PHC showed the strongest effect in the dose-and time-dependent manner; PHC could induce B16 cells dendritic growth 48 h after the treatment, and the morphological changes were typically differentiated; PHC also increased the melanin production and the activity of tyrosinase. There was a significant difference compared to the control group (P < 0.05). After treated by PHC for 6, 12, and 24 h, the expression levels of tyrosinase mRNA, tyrosinase 1 mRNA, and tyrosinase 2 mRNA were significantly increased (P < 0.01) in a time-dependent manner. Conclusion: PHC could inhibit the proliferation of B16 cells and the mechanism is related to the differentiation of B16 cells.

Objective To study the method and effect of superomedial pedicled mammaplasty based on Würinger's horizontal septum.Methods During August 2014 to August 2015,25 cases of cromastia were treated with septum-based mammaplasties with a superomedial pedicled and vertical incision.Based on the preoperation design,the superomedial pedicle was deepithelialized with preservation of the dermis and the subdermal plexus.The pedicle was dissected as described,the excess tissue was removed with the prepectoral fascia preserved.After remodeling the shape of mammary gland,the incision was sutured.Results There was no hematoma,no fat liquefaction and no partial or complete NAC necrosis.The pre and postoperative NAC sensibilities were comparable in all the patients.The patients were followed up for 6 to 12 months,and all the patients were satisfied with the therapeutic effect and expressed acceptance to the scar.Conclusions The breast reduction based on wellvascularized septum and superomedial pedicle is safe with minor scar,even in large breasts.This technique demonstrates ease of pedicle shaping and breast remodeling.The satisfied effect could be obtained for patients with severe ptosis of breasts just after operations and in future.

Objective To explore liver protective effects of the stems and leaves and roots of Scutellariae Radix from different areas of Gansu Province;To discuss differences in liver protection of the stems and leaves and roots of Scutellariae Radix. Methods All Kunming mice were randomly divided into 11 groups:normal group, model group, positive control group, Scutellariae Radix root groups and Scutellariae Radix stem-leaf groups of Longxi, Minxian, Weiyuan, Zhangxian. The mice received gavage with relevant medicine once a day for 7 days. 2 hours after the last medication, the mice were given peritoneal injection with 0.1%CCl4 and olive oil to establish the models with liver injury. Hepatic index, the activity of serum ALT and AST, the levels of T-AOC in the liver tissue were calculated in 18 hours after establishing the models. Results Compared with the model group, hepatic index, the activity of serum ALT and AST in the mice of Scutellariae Radix root groups from difference producing areas decreased significantly (P<0.05);the levels of T-AOC in the liver tissue was raised notably (P<0.05). Compared with stem-leaf groups from the same producing area, hepatic index, the activity of ALT and AST in the mice of Scutellariae Radix root groups were remarkably lower than these of Scutellariae Radix stem-leaf groups (P<0.05);the levels of T-AOC in the liver tissue was evidently higher than that of Scutellariae Radix stem-leaf group (P<0.05). Conclusion Scutellariae Radix root water decoction can prevent and protect CCl4-induced acute liver injury to a certain degree, while Scutellariae Radix stem-leaf water decoction shows no significant protective effect on acute liver injury.

Objective To analyze the distribution and antimicrobial resistance of pathogens isolated from department of hematology during the past three years.Methods Pathogenic strains isolated from patients hospitalized in a hematology de-partment between January 2011 and December 2013 were collected,antimicrobial susceptibility testing was performed by Kirby-Bauer disk diffusion method or automatic system,antimicrobial susceptibility testing results were judged according to American Clinical and Laboratory Standards Institute 2011, data were analyzed by WHONET 5.6 software. Results A total of 462 clinical isolates were collected in 2011—2013,including 161 gram-positive cocci isolates,279 gram-negative bacilli,and 22 fungi.Of Staphylococcus spp ,detection rate of methicillin-resistant coagulase negative Staphylo-coccus(MRCNS)and methicillin-resistant Staphylococcus aureus (MRSA)was 81.37% and 62.50%respectively.The re-sistant rate of Staphylococcus spp .and Enterococcus spp .to linezolid was 1.69% and 3.57% respectively,resistant rate of Staphylococcus spp .to teicoplanin was 3.39%,vancomycin-resistant gram-positive coccus was not found.Enterobacte-riaceae strains Escherichia coli and Klebsiella pneumoniae were highly susceptible to carbapenems,the sensitivity rates were 97.56%—98.88%;while nonfermentative gram-negative bacilli Pseudomonas aeruginosa and Acinetobacter bauman-nii strains were obviously resistant to carbapenems,the resistance rates were 38.71%—64.00%.Conclusion Antimicrobial resistance of major pathogenic strains from hematology department is high,antimicrobial agents should be used according to pathogenic distribution characteristics and antimicrobial susceptibility testing results,healthcare-associated infection control should be strengthened to reduce antimicrobial resistant rate.

Objective To study the effect of interventional treatment on the expression of PgP and GST ? in different histopathological types of primary lung carcinoma. Methods One hundred and eighteen cases of histopathologically verified primary lung carcinoma were studied. SCLC was found in 26 cases and NSCLC in 92 cases. The non chemotherapy group had 50 cases, and the interventional treatment group had 68 cases. PgP and GST ? were examined in all specimens with 2 step immunohistochemistry. Results The positive expression rates of PgP and GST ? were 32.0% and 34.0% in non chemotherapy group, respectively, 75.0% and 78.6% in interventional treatment with non embolization group, respectively, and 50.0% and 52.5% in interventional treatment with embolization group, respectively. The positive expression rates of PgP and GST ? had significant difference between non chemotherapy group and interventional treatment with non embolization group( P 0.05). There was a tendency of positive correlation between differentiated degree of carcinoma and the expression of PgP and GST ? in NSCLC. Condusion To detect PgP and GST ? in carcinoma tissue is important and has the instructive significance for chemotherapy of lung carcinoma. The positive rate of multidrug resistant gene is obviously increased in the primary lung carcinoma with bronchial arterial chemotherapy. The inducement to multidrug resistance gene in bronchial arterial embolization with the emulsifying agent of oil anticarcinogen was lower.

Objective:To study the characteristic of hearing loss in essential hypertension. Method:Sixty-eight cases (136 ears) of patients with essential hypertension were divided into two groups, i. e. group A, patients without retinal alteriosclerosis (35cases, 70 ears ) and group B, patients with retinal alterioselerosis (33cases, 66 ears ). The control group consisted of 30 cases (60 ears) of the same sex and same age. AH people were measured by pure tone audiometry and distortion product otoacoustic emissions. Result:The pure tone thresholds measured in group B hypertensive patients were significantly higher than in the normal controls (P<0. 05) , whereas pure tone thresholds in group A were not different from those in the control group(P>0. 05). From 1000 to 8 000 Hz , DPOAE amplitudes of hypertensive group A and group B were lower than that in control group(P<0. 01). Only on the frequency points of 4 000 Hz , the detection rate of DPOAE in group B was lower than that in control group (P<0. 05). Conclusion: Hypertension is a risk factor of degeneration of the hearing apparatus. The hearing function of hypertensive patients has probability to be impaired, although they do not feel hearing loss.

Objective To assess the effect of EDFS40 on the structure and vasomotoricity of rabbit carotids in cryopreservation. Methods The fresh carotids, taken from rabbits under aseptic conditions, were preserved for 14 days in liquid nitrogen by vitrification or slow freezing method with both fresh and thawed carotids were observed by visual examination, optical and electron microscopes, respectively. Their endothelium-independent vasoconstriction and vasodilatation were detected in organ bath. Results The morphological changes of the EDFS40-vitrified arteries were similar to those of the fresh carotids under macrography, optical and electron microscopes. And there was no obvious difference in contraction to KCl between the cryopreservation group and the slow-freezing group (P>0.05). The contractile and relaxant responses of vitrified arteries to NE or SNP were significantly better than those of the frozen arteries (P<0.05); both, however, were still inferior to the vasomotoricity of the fresh arteries (P<0.05). Conclusion The rabbit carotids vitrified with EDFS40 can have their structure and vasomotor function maintained without obvious damage, which may indicate the usage of EDFS40 in vitrification of blood vessels.

Objective To investigate the surgical strategy of recurrent rectuvaginal fistula. Methods Retrospectivly analyzed the clinical data of 13 patients with recurrent rectovaginal fistula from December 2001 to December 2008. The etiopathogenisis and the ways of treatment were analyzed. Results All the 13 patients with a transverse colostomy surgical repaired with Mason operation in the secondary intention,and recepted closure of colostomy in the third intention for treatment, all patients were cured. No recurrent fistula was identified with postoperative follow-up 4 - 84 months. Conclusions After recurrent rectovaginal fistula with multiple surgical treatment,the blood supply of local tissue is poor,the scar is serious. Selecting the appropriate timing of operation, adequate preoperative preparation, the application of transverse colostomy and Mason operation in different period could significantly enhance the successful operation rate.