Objective To construct the recombinant plasmid that highly expressed human subcellular PTEN,in order to provide a basis for further study on its anti-tumor effect. Methods PTEN cDNA was amplified by RT-PCR based on mRNA of placenta.The PCR product was ligated into T-vector,and transfected into E.coli;the obtained T-PTEN plasmid was identified with restrictive digestion and sequencing.PCR was used to incorporte nuclear signal of localization(NSL) into PTEN when T-PTEN was used as template.Then the PCR product was ligated into T-vector,and transfected into E.coli,and T-NSL-PTEN plasmid was obtained.pcDNA3.1 and T-NSL-PTEN were ligated after digested with EcoRⅠand BamHⅠ,and transfected into E.coli,the recombinant vector pcDNA3.1-NSL-PTEN was obtained,and identified with digestion and sequcncing.Results The recombinant expression vector DUM-PTEN and PUM-NSL PTEN were identified by restrictive digestion and DNA sequencing.As expected,by EcoRⅠ and BamHⅠ digestion,it showed the band of 1 200 bp.The sequencing result showed the NSL was incorporated successfully.The recombinant pcDNA3.1-PTEN was obtained with 1 200 bp,the sequencing result showed that its sequence was same as target gene;the recombinant pcDNA3.1-NSL-PTEN was comfirmed by restrictive digestion and sequencing,and the NSL was incorporated successfully. Conclusion The recombinant expression plasmid pcDNA3.1-NSL-PTEN is constructed successfully which can highly express human subcellular PTEN.

We aimed to make comprehensive cancer care guidelines of China according to American guidelines of National Comprehensive Cancer Network,Ametican Society of Clinical Oncology and Oncology Nursing Society,and to guide clinical nurses to study and draw lessons from foreign latest guidelines,and combined with the actual situation in our country.The results of study and application were advantageous to the nursing profession,the security and interests of the patients.We should develop comprehensive cancer care guidelines of our country,eventually make nursing practice guide system of China constantly improve and perfect.

Objective To observe and compare the incidence rate of hemolysis while venous blood specimen collecting with two types of venous indwelling needle. To summarize the attentions during blood samples collecting by venous indwelling needle. Methods From January 2014 to January 2015, a total of 400 patients in resuscitation room of emergency department were divided into control group and experimental group by random digits table method with 200 cases each. Type Y indwelling needle was used in control group, while straight type indwelling needle was used in experimental group. Both two types of indwelling needle were required to be connected with the disposable venous blood taking needle joint for blood sample collection. The incidence rate of hemolysis was observed. Results The incidence rate of hemolysis in experimental group was lower than that in control group: 8.0%(16/200) vs. 39.5%(79/200),and there was significant difference, χ2=34.317, P < 0.01. Conclusions The incidence rate of hemolysis during venous blood specimen collecting with straight type indwelling needle is significantly lower than that with Y type indwelling needle. Straight type indwelling needle is worthy of clinical use.

Objective To investigate evaluation role of IP-10 level in urine of kidney transplant recipients when using rabbit anti-human T-lymphocyte immunoglobulin to treat acute cellular rejection.Methods A total of 40 patients who underwent renal transplantation and had been diagnosed as acute cellular rejection according to the results of histopathological examination were randomly divided them into IP-10 group (n =20) and serum creatinine group (Scr group,n =20).Urinary IP-10 and Scr levels were measured in time and patients then were treated with ATG,of which the doses and duration were adjusted according to IP-10 or Scr levels.We compared the total and daily ATG dosages,ATG administration period,side effects of ATG such as incidence of severe platelet and neutropenia,acute rejection during first 3 months and infection rates during first 1 year.Result The number of ATG duration is 5.35 ± 1.93 for IP-10 group versus 6.70 ± 1.75 for Scr group.We used a daily dose of 2.50 ± 0.57 mg/(kg·d) for IP-10 group and 2.77 ± 0.74 mg/(kg· d) for Scr group,a total dose of 13.40 ± 6.59 mg/kg for IP-10 group and 18.25 ± 7.35 mg/kg for Scr group.There was significance between the two group in above three outcomes (P ＜ 0.05).There was no significance in incidences of severe thrombocytopenia and neutropenia,incidences of acute rejection during first 3 months,incidences of infection during first 1 year between the two group (P ＞ 0.05).Conclusion Urine IP-10 test is effective and reliable indicators which can guide ATG usage in patients with acute rejection and reduce the ATG cost.

Objective To explore the characteristics and effects of interventional therapy of transplant renal artery rupture of donation after citizens death (DCD).Method Among 28 cases of DCD renal transplantations (from February 2012 to December 2013),the transplant renal artery rupture occurred in 4 cases.Vascular complications were treated with the guide wires to place stents in the pseudoaneurysms or bleeding period.Result Pseudoaneurysms occurred in 2 cases,and they were successfully discharged after interventional treatment.In the rest two patients,the artery residual ruptured and bled after the nephrectomy,and they recovered after interventional treatment to stop bleeding.Conclusion For kidney transplant recipients,the DCD postoperative infection is risky.Some transplant kidneys have local infection and erosion of renal artery,which causes arterial hemorrhage.The interventional treatment of transplant renal artery pseudoaneurysms and rupture bleeding has the advantages of small trauma and instant effect,and can be used as an alternative treatment of open surgery.

Multimedia computing technology teaching is a new kind teaching method. It conquers many shortcomings such as poor video, poor expression in traditional teaching of microbiology. But it can also strangle the improvising creation of different teachers in teaching and then lead it to be typical " computer teaching" . So we should obey the teaching discipline in making multimedia computing technology courseware. It can lead many sections part from the class teaching if just emphasize the full use of video and audio. We should make the purpose obviously, and make it as easy as possible in teaching, and we should also explore the fixed regularity to make the teaching and studying integrated perfectly.

OBJECTIVETo study the effect of Hepatitis B virus X (HBx) gene transfection on expression of human telomerase reverse transcriptase (hTERT) mRNA in human bile duct carcinoma cell lines QBC939 and to elucidate the significance of cis-activation of hTERT mRNA by HBx gene on the carcinogenesis of bile duct.

METHODSQBC939 were cultured in vitro and co-transfected with eukaryotic expression vector containing the HBx coding region and cloning vector containing enhanced green fluorescent protein (EGFP) coding sequence using liposome-mediated gene transduction technique. Thirty six hours after transfection, EGFP expression, the indicator of successful transfection in cells, was determined. Flow cytometry was applied to determine the transfection efficiency. Cells were harvested and total RNA was extracted with TRI(ZOL) Reagent. The expression of hTERT mRNA in QBC939 was assayed by Reverse Transcription Polymerase Chain Reaction. The expression of HBx protein in QBC939 was detected by immunocytochemistry staining and western blotting.

RESULTSThe transfection efficiency was 29.6% for both HBx expression vector and vector control group. The expression of hTERT mRNA was significantly increased when transfected with HBx expression vector than that transfected with OPTI-MEM medium and vector only. The expression of HBx protein could only be found in the cells when transfected with HBx expression vector by immunocytochemistry staining and western blotting.

CONCLUSIONHBx gene transfection may up-regulate the transcriptional expression of hTERT mRNA in bile duct carcinoma cells. The cis-activation of hTERT gene by HBx gene is primary mechanism for carcinogenesis of biliary epithelia after HBV infection.

Bile Duct Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; metabolism ; Trans-Activators ; genetics ; Transfection

OBJECTIVETo explore the efficacy and feasibility of dog days moxibustion plaster therapy in treatment of allergic rhinitis of different patterns/syndromes.

METHODSAllergic rhinitis of lung deficiency and invasion of cold, spleen qi deficiency and kidney yang deficiency, 56 patients for each pattern/syndrome were randomized into a plaster therapy group and a nasal spray group, 28 cases in each one. In the plaster therapy group, according to the pattern/syndrome differentiation, with literature retrieval method, 3 acupoints of high frequency utility in clinic were selected as one group in acupoint plaster therapy. For lung deficiency and invasion of cold pattern/syndrome, Feishu (BL 13), Fengmen (BL 12) and Hegu (LI 4) were selected. For spleen qi deficiency pattern/syndrome, Pishu (BL 21), Zusanli (ST 36) and Dazhui (GV 14) were selected. For kidney yang deficiency pattern/ syndrome, Shenshu (BL 23), Dingchuan (EX-B 1) and Bailao (EX-HN 15) were selected. Separately, on July 13, 2013, July 23, 2013, August 2, 2013 and August 12, 2013, the aucpoint plaster therapy was applied, 2 to 4 h (1 to 2 h for children) each time. In the nasal spray group, beclometasone dipropionate aqueous nasal spray, 2 presses one nostril each time, 2 to 3 times a day, continuously for 4 weeks. The symptom score and efficacy were compared before and after treatment in the patients of the two groups.

RESULTSThe symptom scores of 3 patterns/syndromes were all apparently improved after treatment as compared with those before treatment in the patients of the two groups (all P<0.05), and the result in the plaster therapy group was better than that of the nasal spray group (P<0.05, P<0.01). For lung deficiency and invasion of cold pattern/syndrome, the total effective rate was 87.3% (20/24) in the plaster therapy group, better than 84.6% (22/26) in the nasal spray group (P<0.05). For spleen qi deficiency pattern/syndrome, the total effective rate was 83.3% (20/24) in the plaster therapy group, obviously better than 76.9% (22/26) in the nasal spray group (P<0.05). For kidney yang deficiency pattern/syndrome, the total effective rate was 79.2% (19/24) in the plaster therapy group, better than 76.9% (22/26) in the nasal spray group (P<0.05).

CONCLUSIONThe dog days moxibustion plaster therapy achieves definite efficacy on allergic rhinitis at the acupoints selected based on the differentiation of different patterns/syndromes and the efficacy is better than beclometasone dipropionate aqueous nasal spray.

Acupuncture Points ; Administration, Cutaneous ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Child ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Rhinitis, Allergic ; drug therapy ; therapy ; Yang Deficiency ; drug therapy ; Young Adult

BACKGROUNDThe goal of total knee arthroplasty (TKA) is to restore knee kinematics. Knee prosthesis design plays a very important role in successful restoration. Here, kinematics models of normal and prosthetic knees were created and validated using previously published data.

METHODSComputed tomography and magnetic resonance imaging scans of a healthy, anticorrosive female cadaver were used to establish a model of the entire lower limbs, including the femur, tibia, patella, fibula, distal femur cartilage, and medial and lateral menisci, as well as the anterior cruciate, posterior cruciate, medial collateral, and lateral collateral ligaments. The data from the three-dimensional models of the normal knee joint and a posterior-stabilized (PS) knee prosthesis were imported into finite element analysis software to create the final kinematic model of the TKA prosthesis, which was then validated by comparison with a previous study. The displacement of the medial/lateral femur and the internal rotation angle of the tibia were analyzed during 0-135° flexion.

RESULTSBoth the output data trends and the measured values derived from the normal knee's kinematics model were very close to the results reported in a previous in vivo study, suggesting that this model can be used for further analyses. The PS knee prosthesis underwent an abnormal forward displacement compared with the normal knee and has insufficient, or insufficiently aggressive, "rollback" compared with the lateral femur of the normal knee. In addition, a certain degree of reverse rotation occurs during flexion of the PS knee prosthesis.

CONCLUSIONSThere were still several differences between the kinematics of the PS knee prosthesis and a normal knee, suggesting room for improving the design of the PS knee prosthesis. The abnormal kinematics during early flexion shows that the design of the articular surface played a vital role in improving the kinematics of the PS knee prosthesis.

Adult ; Arthroplasty, Replacement, Knee ; methods ; Biomechanical Phenomena ; Female ; Humans ; Knee Prosthesis

Objective To observe the effect of febuxostat on epithelial-to-mesenchymal transition (EMT) of kidney tubules and the levels of serum IL-6 nad transforming growth factor (TGF) β1 in hyperuricemic rats.Methods Forty male SD rats were divided into 4 groups:normal control group (NC group),oteracil potassium group (OP group),oteracil potassium with febuxostat group (OF group) and oteracil potassium with benzbromarone group (OB group).Each group had 10 rats and balanced in body weights.To induce hyperuricemia,rats were given oteracil potassium by gastric garage once a day for eight weeks.Rats in OF group and OB group were given either febuxostat or benbromarone starting with oteracil potassium,and rats in NC group was given saline only.Blood samples were taken before,and at the end of 4 and 8 weeks of the treatments and serum uric acid,creatinine,blood usea nitrogen (BUN),IL-6 and TGFβ1 contents were measured at each time point.Renal pathological changes were observed via HE and Masson staining,and the expression of α-SMA and E-cadherin were detected by immunohistochemistry.Results Compared with those in NC group,the levels of serum uric acid,creatinine,BUN,IL-6 and TGFβ1 in the another three groups were increased significantly (all P ＜ 0.01).However,the IL-6 and TGFβ1 contents in OF group were much lower than those in OP group (P ＜0.01).HE and Masson staining showed that OF group had less damage and tubulointerstitial fibrosis than OP group and OB group (P ＜0.01).Moreover,the expression of α-SMA was significantly down-regulated (P ＜ 0.01) and that of E-cadherin was significantly up-regulated in OF group compared with those in OP group.Conclusion Febuxostat treatment significantly inhibited EMT and reduced the levels of IL-6 and TGFβ1 in hyperuricemia rats.