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 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 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 investigate the effect of chronic stress on the expression of uncoupling protein 4 (UCP4) and Bcl-2 protein in hippocampal neurons of rat depression model.Methods Rat depression models were established by chronic unpredicted mild stress.All rats were randomly assigned to 2 groups:control group and model group.Flow cytometry was applied to detect the apoptosis rate and mitochondrial membrane potential.The level of LDH was measured by enzymes labelling instrument.The number of neurons was measured by immunohistochemistry.The expression of UCP4 and Bcl-2 protein was measured by Western blotting.Results After chronic stress,the apoptosis rate((4.35±0.19) ％)and LDH activity ((445.50±91.70) U/mg) in hippocampal tissue in the model group was significantly higher than the control group((0.34±0.06) ％,(167.20±63.40)U/mg).Compared to control group,the number of hippocampal neurons ((72.50±4.25) vs (45.30±2.54)) and the mitochon drial membrane potential decreased in the model group.The expressions of UCP4 and Bcl-2 protein in hippocampal tissue were significantly lower than the control group.Conclusion Chronic unpredicted mild stress can lead to apoptosis in rat hippocampal neurons,which is related with decline of mitochondrial membrane potential and low expression of UCP4 and Bcl-2 protein.

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

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.

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