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.

OBJECTIVETo investigate the influence of male age on the pregnancy outcomes of in vitro fertilization-embryo transfer (IVF-ET).

METHODSWe retrospectively analyzed 7,533 cycles of IVF-ET performed between January 1, 2009 and October 31, 2013. We divided the samples into three groups according to the female age (< 30, 30-34, and 35-38 yr), each again subdivided into six groups based on the male age (< 30, 30-32, 33-35, 36-38, 39-41, and ≥ 42 yr). We compared the rates of implantation, pregnancy, miscarriage, and live birth among different age groups.

RESULTSThere were no statistically significant differences in basal E2, FSH, endometrium thickness on the day of hCG administration, number of oocytes retrieved, and days of embryo transfer among different male age groups (P > 0.05). The implantation rate showed an age-dependent decrease in the < 30, 30-32, 33-35, 36-38, 39-41, and ≥ 42 yr male groups, 41.1, 42.0, 39.5, 31.3, 40.7, and 48.6% among the women aged < 30 years (P < 0.05), 40.3, 36.4, 35.1, 35.3, 29.4, and 37.3% among the women aged 30-34 years (P < 0.05), and 48.2, 17.8, 25.3, 23.5, 22.1, and 23.8% among the women aged 35-38 years (P < 0.05). The miscarriage rate was significantly higher in the ≥ 39 yr than in the 30-32 and 33-35 yr male age groups among the women aged 30-34 years (P < 0.05), but showed no remarkable differences among the other male age groups in the women aged < 30 and 35-38 years (P > 0.05). No statistically significant differences were observed in the rates of pregnancy and live birth among different male age groups (P > 0.05).

CONCLUSIONMale age has some influence on the rates of implantation and miscarriage but not on the rates of pregnancy and live birth in IVF-ET.

Abortion, Spontaneous ; epidemiology ; Adult ; Age Factors ; Embryo Implantation ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Male ; Oocyte Retrieval ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies ; Sex Factors

Adolescent ; Adult ; Aged ; Cholesteatoma, Middle Ear ; congenital ; pathology ; Female ; Humans ; Male ; Melanocytes ; Middle Aged ; Young Adult

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 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 evaluate the clinical efficacy and security of retroperitoneal laparoscopic ureterolithotomy(RLUL) for impacted ureteral calculi after radical cystectomy and ileal conduit.Methods 5 patients with unilateral impacted ureteral calculi after radical cystectomy and ileal conduit received RLUL were selected,and a retrospective study was performed for manner,duration of surgery,complications and length of stay and other indicators,and the treatment effect was evaluated.Results All procedures were successful and the mean operation time was (82.0±27.7)min,the extubation time was (5.4±1.1)d,hospitalization time was (10.0±2.9)d.Postoperative follow up for 6-48 months,there were no obvious complications.Conclusion The RLUL showed satisfactory availability and security for management ureteral calculi after radical cystectomy and ileal conduit.