Objective To construct GST/HIF1α fusion protein expression vector and induce its expression in Escherichia coli (E.coli). Methods The coding sequence of hypoxia inducible factor-la (HIF-la) and its deletion fragments were amplified from the plasmid pcD-NA3.1-HIF-la by PCR and inserted into pGEX-4T-2 by BamHI and Not I. The positive recombinanls were identified by restriction endonu-clease digestion and DNA sequencing. Then they were transformed into E.coli BL21, induced by IPTG and identified by SDS-PAGE and Western blot. Results The prokaryotic expression plasmid pGEX-4T-2-HIF-lα and its deletion mutants were successfully constructed and confirmed by enzyme digestion and sequencing. The desired CST/HIF-1α fusion proteins were expressed and confirmed by Western blot. Conclusion The prokaryotic expression plasmid of HIF-la and its deletion mutants were successfully constructed and the expression of fu-sion proteins was confirmed. This study provides the basis for the further research on purifying HIF-la protein and the biological function of HIF-lα.

Objective To monitor the activities of orbicularis oculi muscle by analyzing the electroneurogram (ENG) of its innervating nerve. Methods The ENG was recorded in vivo by a cuff electrode implanted around a dog’s zygomatic branch of facial nerve. The differences between the ENG during relaxing period of the orbicularis oculi muscle and during the constricting period were analyzed to explore a way to monitor the muscle activities. Results The ENG signals indicating occurrences of eye blinking events were extracted in the study. Time-frequency analysis showed that there was no ENG difference when the orbicularis oculi muscle was in relaxing periods no mater whether it was in the nature eye-opening period or in the interval of eye blinking. The ENG of orbicularis oculi muscle in constricting period was different from the ENG in relaxing period at amplitude and frequency. Conclusion It is feasible to monitor the orbicularis oculi muscle activities by ENG analysis.

ObjectiveTo observe the effect of Integrated Chinese-western three-stage rehabilitation on activities of daily living (ADL) in stroke patients. Methods60 stroke patients were randomly divided into the trial group and control group with 30 cases in each group. Patients of control group were treated with routine neurological interventions and three-stage rehabilitation, while those of trial group were administered resuscitating acupuncture therapy, scalp and body acupuncture in addition. All the patients were assessed with modified Barthel index (MBI) at the beginning, 1 month, 3 months, 6 months respectively after stroke. ResultsThere was no significant difference in ADL between the trial and control groups at the beginning, and both improved at the following period (P<0.01). At each of the following period, either the ADL scores (P<0.05) or the increase of the ADL scores (P<0.05) of the trial group were significant higher than those of the control group. ConclusionIntegrated Chinese-western three-stage rehabilitation could improve the ADL further for the stroke patients.

A strain ph 16 , that could effectively degrade phenol,was isolated from sewer sludge of printing and dyeing plant. The preliminary identification sugg ested that the strain belongs to Micrococcus sp. The strain could resist to phenol up to 1.5 g/L. The efficient biodegradation of phenol occurred when th e strain was cultured in the medium (pH 7.0) containing 1.0 g/L phenol under 35℃, wh er e the highest degradation rate reach 99.6% after 36 hours. This strain, when t re ated with some heavy metal ions such as Hg +、Co 2+ and Ag 2+ , showe d the significant inhibition of phenol degradation by 74.2%～100%. The kinetic s of phenol degradation during culture of the strain was also explored.

In order to develop an anti-FMDV A Type monoclonal antibo by (mAb),BABL/c mice were immunized with FMDV A type.Monoclonal antibodies (mAbs) 7B11 and 8H4 against Foot-and-mouth disease virus (FMDV) serotype A were produced by fusing SP2/O myeloma cells with splenocyte from the mouse immunized with A/AV88.The microneutralization titer of the mAbs 7B11 and 8H4 were 1024 and 512,respectively.Both mAbs contain kappa light chains,the mAbs were IgG1.In order to define the mAbs binding epitopes,the reactivity of these mAbs against A Type FMDV,were examined using indirect ELISA,the result showed that both mAbs reacted with A Type FMDV.These mAbs may be used for further vaccine studies,diagnostic methods,prophylaxis,etiological and immunological research on FMDV.Characterization of these ncindicated that prepared anti-FMDV A mAbs had no cross-reactivity with Swine Vesicular Disease (SVD) or FMDV O,Asial and C Type antigens.Their titers in abdomen liquor were 1:5×106 and 1:2×106,respectively.7B11 was found to be of subtype IgG1,8H4 was classified as IgG2b subtype.The mAbs prepared in this study,are specific for detection of FMDV serotype A,and is potentially useful for pen-side diagnosis.

Objective To evaluate the surgical treatment of the acute posterolateral complex(PLC)injuries of knee joint and then observe the clinical outcome.Methods Twelve cases(12 knees)of acute PLC injuries were treated from May 2006 to October 2008.Patients' age ranged from 23 to 47 years old,average 31 years.There were 9 males and 3 females.Rebuild the anterior cruciate ligament(ACL)and posterior cruciate ligament(PCL)under arthroscope and then,locally suture the PLC injuries sites on those patients with PLC avulsion fraction.If there is PLC rupture,then locally suture the injury sites plus PLC reconstruction.Knee functions were evaluated by IKDC and Lysholm score.Results All patients were followed up for 12-18 months(mean,13.3 months).The preoperative range of motion was 118.00°±6.77°,which was 130.75°±3.05° after surgery.KT-1000 arthrometer measurement showed that the average posterior translation improved from(14.85+1.83)mm preoperatively to(4.18±1.88)mm postoperatively.Seven cases were normal(A grade),3 cases were nearly normal(B grade),1 abnormal(C grade),and 1 severely abnormal(D grade)according to IKDC standard.The preoperative Lysholm joint function score was 35-44,average 38.83 ±3.16,which was 79-91,average 84.92±3.73 after surgery.Conclusion To those acute PLC injuries with avulsion at the ligament extremities,locally suture should be taken.But for those with PLC rupture at the mid part of ligament,locally suture the injury sites plus PLC reconstruction helps get satisfactory outcome.

Adult ; Female ; Humans ; Immunohistochemistry ; methods ; Male ; Meningeal Neoplasms ; immunology ; ultrastructure ; Meningioma ; immunology ; ultrastructure ; Microscopy, Electron ; Middle Aged ; Paraganglioma ; physiopathology

Objective To investigate circulating level of growth differentiation factor-15 (GDF-15) in patients with non-valvular atrial fibrillation (AF). Methods We enrolled 104 consecutive patients with AF (67 are paroxymal AF and 37 are persistent AF) that presented to our hospital between August 2012 and June 2013. Subjects without AF (n=67) was used as controls who were matched for sex, age and atherosclerotic risk factors enrolled in the same period. Serum GDF-15 con-centration was detected by enzyme linked immunosorbent assay (ELISA). Echocardiography and laboratory parameters were compared among three groups. The correlation of GDF- 15 and hypersensitive c-reactive protein (hs-CRP) as well as the risk factors of development of AF were also analyzed. Results No significant difference in age, history of hypertension, coro-nary heart disease, and stroke was found among three groups (P>0.05). GDF-15 had significant difference between pa-tients with paroxysmal AF and controls [(1 473.14±628.52)μg/L vs (1 233.592±262.76)μg/L, P=0.012]. GDF-15 level was correlated with hs-CRP (rs=0.172,P=0.044). Multivariable analysis demonstrated that high GDF-15 is a risk factor of devel-opment of paroxysmal AF/AF (OR:1.002, 95%CI:1.000-1.003, P<0.05). Conclusion Patients with AF have a higher se-rum level of GDF-15 compared with controls and GDF-15 level is a risk factor of development of paroxysmal AF/AF.

Objective:To assess the efficacy of trastuzumab combined with neoadjuvant chemotherapy in the treatment of human epidermal growth factor receptor-2 (Her-2)positive breast cancer patients,and to explore its influencing factors of prognosis of breast cancer.Methods:The clinical materials of 112 Her-2 positive breast cancer patients were collected and they were divided into combined treatment group (trastuzumab + neoadj uvant chemotherapy)and single chemotherapy group (without trastuzumab ). SPSS 1 9.0 software was employed to calculate and analyze their clinical characteristics.The survival rate and prognosis were analyzed by Kaplan-Meier method,Log-rank test, and Cox regression. Results:Among the 112 Her-2 positive breast cancer patients, 23 cases were treated with trastuzumab,89 cases were treated without trastuzumab.The disease-free survival (DFS)in combined treatment group and single chemotherapy group had significant difference (P=0.012).And there was no significant difference of overall survival (OS)between two groups (P=0.064).The HR negative group had 18 (32.7%)patients with 5-level of Miller and Payne (MP)classification which was higher than HR positive group (5/46,10.9%)(P=0.009).In univariate analysis,the tumor size,node status at diagnosis and node status after operation were the influencing factors of DFS in the Her-2 positive breast cancer patients;the tumor size,node status at diagnosis,MP classification and node status after operation were the influencing factors of OS in the Her-2 positive breast cancer patients.The result of multivariate analysis indicated that the tumor size, node status at diagnosis,and with or without radiotherapy were the independent influencing factors of DFS and OS in the Her-2 positive breast cancer patients. Conclusion:The breast lesions in the Her-2 positive breast cancer patients with negative HR are relieved after treated with trastuzumab combined with neoadjuvant chemotherapy. The combination of trastuzumab and neoadj uvant chemotherapy can significantly improve the DFS and has a positive influence in the prognosis of Her-2 positive breast cancer patients.

Objective: To observe the effect of acupoint massage plus Vitalstim electrical stimulation on deglutition function and surface electromyography (SEMG) of deglutition muscle groups. Methods: A total of 60 patients with deglutition disorder after stroke were selected and divided into an electrical stimulation group, a massage group and an integrated group according to the random number table method, with 20 cases in each group. Patients in these three groups were given the same routine rehabilitation training for deglutition. In addition, patients in the electrical stimulation group were given extra Vitalstim electrical stimulation, patients in the massage group were given extra acupoint massage on the head, face and neck, and patients in the integrated group were given extra acupoint massage plus Vitalstim electrical stimulation. Fujishima Ichiro food intake level scale (FILS) was scored before and after treatment. The swallowing duration and maximal amplitude of masseter muscle in SEMG were evaluated before and after treatment. Results: After treatment, the FILS score and the maximal amplitude of recruitment potential generated by muscular contraction of masseter muscle group in the three groups were higher than those before treatment (all P<0.05), and the swallowing duration of masseter muscle group was shortened compared with that in the same group before treatment (all P<0.05). After treatment, the FILS score in the integrated group was higher than that in the electrical stimulation group and the massage group (both P<0.05). The swallowing duration of masseter muscle group measured by SEMG was lower than that in the electrical stimulation group and the massage group (both P<0.05), while the maximal amplitude was higher than that of the electrical stimulation group and the massage group (P<0.05). After treatment, there were no significant differences in the FILS score, swallowing duration and maximal amplitude of masseter muscle group between the electrical stimulation group and the massage group (all P>0.05). Conclusion: Both acupoint massage and electrical stimulation can improve the deglutition function in patients with deglutition disorder after stroke, and improve the coordination and flexibility of masseter muscle. The integration of the two is more effective.