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.

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

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 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 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 discuss the role of clinical pharmacists in perioperative drug treatment for a patient after renal transplan-tation. Methods:Clinical pharmacists participated in the drug therapy for a patient after renal transplantation. Pharmaceutical care and health education were performed for the patient in the aspects of immunosuppression, anti-hepatitis B virus, gastric mucosa protection, diarrhea, acute rejection and the other treatment programs. Results:The participation of clinical pharmacists in the clinical treatment for the patient improved therapeutic scheme and promoted therapeutic efficacy. Conclusion:The participation of clinical pharmacists in clinical drug therapy for patients can provide individualized service to improve reasonable, effective and economical drug use.

Objective To compare distribution and antimicrobial resistance of methicillin-sensitive Staphylococcus aureus(MSSA)and methicillin-resistant Staphylococcusaureus(MRSA)in hospitalized children,and provide refer-ence for empirical use of antimicrobial agents.Methods Isolation and clinical data of Staphylococcus aureus (S. aureus)from hospitalized children in a hospital during 2011-2015 were analyzed retrospectively,distribution and antimicrobial resistance between MSSA and MRSA were compared.Results A total of 919 strains of S.aureus were isolated,632(68.77% )of which were MSSA,287(31.23%)were MRSA.65.03% of MSSA infection and 64.11% of MRSA infection were in children aged 29 day-1 year old.80.38% of MSSA and 79.09% of MRSA were isolated from sputum specimen.MSSA and MRSA were mainly distributed in department of pediatric respiratory medicine(50.73%,45.89% respectively)and department of pediatric neurology(22.98%,26.84% respectively). Resistance rates of MSSA to antimicrobial agents were<20.00% except penicillin and erythromycin;resistance rates of MRSA to penicillin,oxacillin,erythromycin,and clindamycin were all>40.00%;resistance rates of MR-SA to tetracycline,erythromycin,clindamycin,levofloxacin,ciprofloxacin,moxifloxacin,nitrofurantoin,and ri-fampin were all higher than MSSA.Conclusion MSSA is main S.aureus isolated from hospitalized children,in-fants under 1 year of age are the main population,the main distribution departments of MSSA and MRSA from re-spiratory tract specimen are similar,antimicrobial resistance of MRSA is generally higher than that of MSSA.