To reduce the risk of 3′-terminal mismatch between primers and template and increase the sensitivity of polymerase chain reaction (PCR) in the detection of variable region of DNA. Methods: A pair of special primer(WU,WD) was designed to amplify a fragment of HBV DNA P gene by PCR. Other 2 similar pairs of primer (MU1, MD1, MU2, MD2) were obtained by knocking off 1 or 2 bases at the 3′-terminal of WU and WD. (1) Special primers (WU, WD) and degeneracy primers(WU, WD, MU1, MU2, MD1, MD2) were used to amplify 27 samples respectively by PCR under the same condition. The sensitivity of each PCR was compared. (2) Using degeneracy primers, serum HBV DNA was amplified from 4 patients who were resistant to lamivudine. The PCR products were sequenced to evaluate the effect of the 3′-terminal mismatch of primers upon PCR. Results: (1) The sensitivity of special primers and degeneracy primers were 70.4%(19/27) and 85.2%(23/27) respectively (P<0.05). (2) The sequencing analysis of the PCR products suggested that the 3′-terminal mismatch of primers caused false negative in the PCR detection. Conclusion: When amplifying the variable region of DNA, the false negative result can be avoided by using 3′-terminus shifted degeneracy primers.

Objective To evaluate the effect of a novel orthopedic biomaterial,graded composite zireonia(ZrO2)hydroxyapatite(HAP)on activity of rat osteoblast ceils(OB)cultured in vitro. Methods The pure zirconia material was used as control to measure surface roughness of the composite material that was examined by using scanning electron microscope(SEM)and X-ray diffraetometer (XRD).The rat osteoblast cells were cultured on the two materials.Alkali phosphatase(ALP)of the two groups was measured and ELISA was used to detect IL-6 and TGF-?eoncentration of the supematant of OB cells.Tumor growth factor-?(TGF-?)mRNA was detected by RT-PCR.SEM was used to observe OB cells on the two materials.The extract of the composite material was used for a eytotoxicity test to cal- culate the relative proliferation rate(RGR)and classify the toxicity.Results The surface roughness of the gradual composite materials was significantly higher than that of the control materials(P＜0.01). The ALP of the gradual material group was markedly higher than that of the control group at different in- tervals.There was significant difference of the IL-6 and TGF-?concentrations 2-4 days after culture be- tween two groups(P＜0.05,P＜0.01).The mRNA level of TGF-?of the two OB groups also showed marked statistical difference(P＜0.01).The ossification of the OB cells on the composed material was marked after 14 days.The MTT color experiments showed no statistic significance between materials group and negative group,with the toxicity at levelⅠand 0(P＜0.05).Conclusion Graded composite ZrO2 HAP can significantly promote proliferation and differentiation of OB cells cultured in vitro and has good biocompatibility.

Animals ; Brain ; drug effects ; metabolism ; Carbon Dioxide ; metabolism ; Chronic Disease ; Curcumin ; pharmacology ; Disease Models, Animal ; Fas Ligand Protein ; metabolism ; Hypoxia ; metabolism ; Male ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; fas Receptor ; metabolism

OBJECTIVETo explore clinical effects of suturing-assisted locking plate in treating elderly proximal humeral fractures.

METHODSFrom January 2005 to January 2013, 55 elderly patients with three- and four-part fractures of proximal humeral fractures were divided into treatment group and control group. In treatment group, there were 31 patients including 12 males, and 19 females aged from 65 to 85 with an average of (74.00±5.42) years old, and treated with suturing-assisted locking plates; 19 patients were Neer 3-part fractures, and 12 patients were Neer 4-part fractures of proximal humerus; 23 patients were suffered from low-energy injuries and 8 patients were caused by high-energy injuries. In control group, there were 24 patients including 7 males, and 17 females aged from 65 to 85 with an average of (72.79±5.34) years old, and treated with locking plates; 16 patients were Neer 3-part fractures, and 8 patients were Neer 4-part fractures of proximal humerus; 17 patients were suffered from low-energy injuries and 7 patients were caused by high-energy injuries. Operative time, blood loss during operation, and bone healing time between two groups were observed and compared. Postoperative Neer scoring were used to evaluate recovery of shoulder joint function.

RESULTSAll patients were followed up from 6 to 24 months with an average of 16.1 months. In treatment group, blood loss was (495.806±143.150) ml, function of Neer scoring was 22.645±2.443, range of action was 18.194±2.613, anatomy was 7.935±1.504 and total score of Neer scoring was 77.161±8.335; while in control group, blood loss was (641.667±169.851) ml, function of Neer scoring was 13.958±1.989, range of action was 13.083±2.165, anatomy was 5.500±1.978 and total score of Neer scoring was 58.792±7.313. There were sigificant difference between two groups in these indexes.

CONCLUSIONSuturing-assisted locking plate for the treatment of proximal humerus fractures in elderly, has advantages of less blood loss, simple fracture reduction and rapid recovery of shoulder joint, and is a effective method.

Aged ; Aged, 80 and over ; Bone Plates ; Case-Control Studies ; Female ; Humans ; Male ; Recovery of Function ; Shoulder Fractures ; physiopathology ; surgery ; Shoulder Joint ; physiopathology ; Sutures

In this study the treatment effect of combined implantation of autologous skin on pig dermis in injured rats was observed. Twenty-one Wistar rats were used, and the wounds were formed by excising a piece of full thickness skin on the back. After the pig dermis was implanted, the autologous skin was grafted on the dermis at 0.7 and 10 days. In the group with perforated pig dermis, the autograft skin was implanted on the day when the pig dermis was implanted. The healing effect was evaluated by measuring wound area, and by observing the growth of the autograft skin. Two weeks after the autograft skin was implanted, the skin securely adhered to the dermis, and the edge of autograft skin expanded clearly. The wound of the autograft skin implanted in the perforation of the dermis healed completely after 3 weeks, but the other 3 groups had remnant small wound. The autograft skin merged with the dermis and its surrounding tissue, but a clear dividing line still existed between autograft skin and dermis after implantation. The area of the implanted dermis and autograft skin varied from 51.8% to 65.9% compared to its original size. The results suggested that the time and the way of autologous skin grafting on xenogenous dermis may influence wound contraction and healing time.
Animals ; Dermis ; transplantation ; Female ; Graft Survival ; Male ; Rats ; Rats, Wistar ; Skin Transplantation ; methods ; Swine ; Swine, Miniature ; Transplantation, Autologous ; Transplantation, Heterologous

Adolescent ; Child ; Female ; Hip Dislocation, Congenital ; diagnostic imaging ; pathology ; surgery ; Humans ; Male ; Osteotomy ; methods ; Pelvic Bones ; surgery ; Radiography

Fermentation and induction conditions for recombinant Escherichia coli expressing Thermobifida fusca cutinase were optimized in flasks and 3L fermenter. Surface modification of poly (ethylene terephthalate) fibers with cutinase was also discussed. The results showed that, cutinase yield reached 128 U/mL by adding 2 g/L inducer lactose and 0.5% glycine. In the fed-batch culture in a 3 L fermenter, the maximum biomass cutinase activity was up to 506 U/mL, which is the highest bacterial cutinase activity reported by far. Recombinant cutinase was used to modify polyester fibers and terephthalic acid substance was detected by using UV analysis. The dyeing and wetting properties of cutinase treated fibers were higher than untreated fibers. Combined utilization of cutinase and Triton X-100 can significantly improve the hydrophilicity of polyester. This is the first report of surface modification on polyester fibers by bacterial cutinase.
Actinomycetales ; enzymology ; genetics ; Carboxylic Ester Hydrolases ; biosynthesis ; chemistry ; genetics ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Polyethylene Glycols ; chemistry ; Polyethylene Terephthalates ; Recombinant Proteins ; biosynthesis ; genetics ; Surface Properties

PURPOSE: The aim of the study was to evaluate the efficacy and safety of combining sorafenib with chemotherapy in patients with human epidermal growth factor receptor 2 (HER2)-negative advanced breast cancer. METHODS: MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, American Society for Clinical Oncology abstracts, and European Society for Medical Oncology abstracts were searched. Randomized clinical trials that compared the efficacy and safety of sorafenib plus chemotherapy in patients with HER2-negative advanced breast cancer with placebo plus chemotherapy were eligible. The endpoints were progression-free survival (PFS), overall survival (OS), time to progression (TTP), duration of response (DOR), overall response rate (ORR), clinical benefits, and adverse effects. The meta-analysis was performed using Review Manager 5.2.6 (The Nordic Cochrane Centre), and the fixed-effect model weighted by the Mantel-Haenszel method was used. When considerable heterogeneity was found (p<0.1), further analysis (subgroup analysis, sensitivity analysis, or random-effect model) was performed to identify the potential cause. The results are expressed as hazard ratios or risk ratios, with their corresponding 95% confidence intervals. RESULTS: The final analysis included four trials comprising 844 patients. The results revealed longer PFS and TTP, and higher ORR and clinical benefit rates in patients receiving sorafenib combined with chemotherapy compared to those receiving chemotherapy and placebo. OS and DOR were similar in the two groups. Meanwhile, the incidence of some adverse effects, including hand-foot skin reaction/hand-foot syndrome, diarrhea, rash, and hypertension, were significantly higher in the sorafenib arm. CONCLUSION: Sorafenib combined with chemotherapy may prolong PFS and TTP. This treatment was associated with manageable toxicities, but frequent dose interruptions and reductions were required.
Arm ; Breast Neoplasms* ; Breast* ; Diarrhea ; Disease-Free Survival ; Drug Therapy* ; Exanthema ; Humans ; Hypertension ; Incidence ; Medical Oncology ; Odds Ratio ; Population Characteristics ; Receptor, Epidermal Growth Factor ; Skin ; Treatment Outcome

Objective To investigate drug resistance genes and epidemic characteristics of β-lactamase carried by carbapenem-resistant Acinetobacter baumannii (CRAB) in the respiratory intensive care unit(RICU) in a hospital.Methods Clinically isolated CRAB from RICU patients in October-December 2015 were collected.Five drug resistance genes (KPC-2,IMP,VIM,NDM-1,OXA-23) were specifically amplified by polymerase chain reaction (PCR),amplified products were performed agarose gel electrophoresis and sequencing analysis,the homology was analyzed with pulsed-field gel electrophoresis (PFGE).Results A total of 22 CRAB strains were isolated in October-December 2015,19 (86.36％) of which were isolated from sputum.The resistance rate of 22 CRAB strains to compound sulfamethoxazole was 59.09 ％,resistance rate to minocycline was 9.09 ％,all were sensitive to polymyxin B,resistance rates to other antimicrobial agents were more than 80％.Three kinds of resistance genes KPC-2,IMP and NDM-1 were not found by PCR amplification,positive rates of VIM and OXA-23 were both 100％.PFGE homology analysis revealed that 22 strains were divided into 13 different types,each type contained 1-5 strains,9 types(69.23％) contained only 1 strain respectively,the other 4 types (30.77％) contained 2-5 strains.A5,A7,and A8;A9,A11,A14,A19 and A22;A4,A10 and A12;A16 and A18 were of the same type respectively.Conclusion The main types of β-lactamase-resistant genes of CRAB in RICU are VIM and OXA-23.Homology analysis shows a small parts are of the same clone strains,which reveals epidemic of a small scale.

OBJECTIVE: To investigate the effect of lidocaine on LPS induced apoptosis of cultured adult rat alveolar Type II (AT-II) cells. METHODS: Cultured cells were exposed to LPS and lidocaine for 24 hours. Apoptosis and necrosis rates of cells were detected by flow cytometry and electron microscope. The activity of lactic dehydrogenase (LDH) was analyzed by using LDH kits. RESULTS: LPS induced the AT-II cell injuries by increasing not only the necrosis and apoptosis rates but also the LDH release of cultured AT-II in vitro. Lidocaine decreased the necrosis and apoptosis rates of AT-II cells. CONCLUSION Lidocaine can directly inhibit the apoptosis and necrosis induced by LPS in cultured AT-II cells.
Animals ; Apoptosis ; drug effects ; Cells, Cultured ; Epithelial Cells ; pathology ; Flow Cytometry ; Lactate Dehydrogenases ; metabolism ; Lidocaine ; pharmacology ; Lipopolysaccharides ; Necrosis ; Protective Agents ; pharmacology ; Pulmonary Alveoli ; pathology ; Rats ; Rats, Sprague-Dawley