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

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.

Objective To determine the level of five trace elements(Fe 2+ ?Cu 2+ ?Zn 2+ ?Ca 2+ ?Mg 2+ )in the spleen and changes of T lymphocyte and its subtype variations in peripheral blood from the rats infected with Toxoplasma gondii . Methods Twenty rats were randomly and equally divided into two groups: control group and experiment group. Each rat in the experiment group received an ip injection of 2 ml normal saline containing 1.5?10 6 tachyzoites of T. gondii . On the 64th day after injection of T.gondii , the changes in T lymphocytes (TL) and their subgroups, the helper T lymphocytes (Th) and the suppressor T lymphocytes(Ts) in the peripheral blood of the rats with T.gondii were determined by the assay of the lymphocytes labeled with intercellular acid ? naphthyl acetate esterase. All the rats were killed and the atomic absorption method were used for detecting the level of trace elements in the spleen tissue. Results The number of TL and Th in experiment group was significantly lower than that of control ( P

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 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 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.