Objective To clone, express and purify human PIF1 protein and analyze its ATPase activity. Methods The PIF1 cDNA was amplified by PCR from HeLa cell cDNA library and inserted to pET24b with histidine tag at its terminus to form pET24b-PIF1 plasmid. The recombinant pET24b-PIF1 plasmid was transformed to RosettaTM 2 (DE3) and the expression of PIF1 protein was monitored by SDS-PAGE analysis. By using fast protein liquid chromatograph (FPLC) system, the PIF1 protein was purified by affinity chromatograph and gel filtration. The ATPase activity of PIF1 was checked by thin layer chromatograph(TLC). Results The PIF1 protein was successfully cloned and expressed in E.coli. Conclusions The purification procedure of PIF1 protein was established using FPLC. The overexpressed and the purified PIF1 helicase has DNA and Mg2+ dependent ATPase activity.

Objective To detect the expression of the tryptase in the plasma,and study the meaning in brain traumatic patients.Methods There were two groups:the brain traumatic group(40 patients)and the control group (20 health people).The content of plasma tryptase was determined by fluorescence enzyme immunoassay..Results The level of plasma tryptase had no statistical significance in control group(2.97 ±1.05)μg/L compared with the brain traumatic group(3.03 ±1.39)μg/L,however there had statistical significance comparing with sever brain traumatic patients(3.84 ±0.52μg/L)(t =3.32,P <0.05).4 cases of death in patients with severe head injury group content of tryptase (5.85 ±1.05)μg/L,which was significantly higher than the group of 16 cases of injury in severe head injury after 2 months still alive with the content of serum tryptase (2.49 ±0.52)μg/L,the difference was statistically significant (t =8.13,P <0.01).Conclusion The plasma tryptase level in sever brain traumatic patients increased significantly,and might be of importance for treatment strategies and prognosis.

Enhanced biological phosphorus removal (EBPR) is widely accepted as one of the most economical and sustainable processes to remove phosphorus from wastewater.Poor performance or complete failure of EBPR processes has been substantially reported because of the proliferation of glycogen-accumulating organisms (GAOs) in the system.This paper presented the GAOs' metabolic mechanism and the impact factors, such as influent substrate, P/C ratio, pH value, temperature and SRT, on competition between GAOs and PAOs to better understand GAOs' characteristics and improve the performance and reliability of EBPR systems.

Objective To study the relationship between chromosomal abnormalities of diffuse large B-cell lymphoma and its survival time.Methods Chromosome preparations were made by using modified method.Karyotypes were analyzed by stain of G-banding. And all patients were treated by chemotherapy. All patients' survival time was calculated.Results Mitotic cells that could be used for analysis were found in 28 cases.5 of 28 karyotypes were normal and 8 cases were polyploid.There were 4 cases with t(14,18)(q32;q21),5 cases with t(3; 14) (q27;q32),2 cases with t(2;3) (p11 ;q27),1 case with t(3 ;22) (q27 ;q11) respectively.There were 2 cases with ectopia between 7 chromosome and other chromosomes and 1 case with ectopia between 17 chromosome and other chromosomes.The survival time of patients with normal karyotype,t(14,18) (q32;q21)or 3q+ was longer than that of other groups.The survival time of group in Ⅰ, Ⅱ stages was longer than that in Ⅲ, Ⅳ stages. Conclusion The treatment, survival time and prognosis could be expected according to chromosomal abnormalities and its relevance to stages in diffuse large B-cell lymphoma.

Objective To compare the advantages and disadvantages of non-irrigation and irrigation in the surgical approach of chronic subdural hematoma (CSDH),thus to provide reference for clinical treatment of CSDH.Methods Clinical data of 102 patients with CSDH were retrospectively analyzed.According to the different operation methods,the patients were divided into the non-irrigation group(52 cases) and the irrigation group(50 cases).The blood loss during the procedure,operative time,length of stay and postoperative complication rate between the two groups were compared,and the causes of postoperative complications were analyzed.Results The blood loss during the procedure,operative time and length of stay in the non-irrigation group were (6.73 ± 1.17) mL,(15.06 ± 2.64) min and (10.74 ± 2.20) d,respectively,which in the irrigation group were (19.52 ± 3.18) mL,(38.54 ± 6.95) min and (10.44 ± 2.07)d,respectively,there were statistically significant differences between the two groups in the blood loss during the procedure and the operative time (t =-27.11,-22.72,all P ＜ 0.05),there was no statistically significant difference between the two groups in the length of stay (t =0.70,P ＞ 0.05).The incidence rates of postoperative complication in the non-irrigation group and irrigation group were 8.00％ and 7.69％,respectively,there was no significant difference between the two groups (x2 =0.003,P ＞ 0.05).Conclusion Each of the two methods has its own advantages and disadvantages in the treatment of CSDH.Compared with burr hole irrigation,burr hole non irrigation has the advantages of less blood loss and shorter operative time.However,burr hole non-irrigation is more likely to suffer serious complication.We should select suitable surgical approach by the specific circumstances of the patients.The causes of postoperative complications of CSDH are varied.In particular,there is an important relationship between the non-standard operation and postoperative complications.

Adult ; Aged ; Breast Neoplasms ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; methods ; Mastectomy, Modified Radical ; Middle Aged ; Ultrasonic Therapy ; methods ; Young Adult

Objective To discuss the diagnosis and treatment of inflammatory granuloma in central nervous system(CNS)to provide reference for clinic.Methods Retrospective data included 8 patients with CNS inflammatory granuloma in Department of Neurosurgery,Shanxi People's Hospital,2012 -2015.We analyzed the imaging features, postoperative symptoms,blood and cerebrospinal fluid changes and prognosis.Results 8 cases all received surgical treatment.All the symptoms were improved,and the CT showed that the lesions were disappeared.All the patients had recovered to normal life and work.Conclusion The diagnosis of CNS inflammatory granuloma is difficult.Clinical manifestations are lack of specificity.The blood and cerebrospinal fluid laboratory examination have no abnormal changes.CT and MRI are the main diagnostic methods.Postoperative pathology is the gold standard for diagnosis.The large lesion,frequent episodes of epilepsy,severe neurological deficits and possibility of brain tumor all should be treated by surgery.

Objective To establish a method for determining oxalate and citrate in urine simultaneously by capillary electrophoresis.The components,the concentration and pH of the buffer solution,the separation voltage and the injection time on theseparation were studied in detail.Methods The separations were carried out using potassium dihydrogen phosphatebuffer ina fused-silica capillary tubeby capillary zone electrophoresis (CZE) and the detection were monitored by UV.24 h-urine samples from patients (n =5) and health control (n =5) were collected from Zhongshan Hospital of Fudan University for systematically validating the method developed.Results The optimized separations were carried out using a 50 mmol/L potassium dihydrogen phosphatebuffer solution (pH 6.5) in a fused-silica capillary tube of 50 cm × 50 μm I.D.Injections were made by using the pressure mode for 10 s at 34 mbar.The detections were monitored by a UV at 200 nm after samples were separated at avohage of 30 kV.Under the seconditions,urinary oxalate and citrate were separated completely within 5 min.The relative standard deviations of migration time and peak area within-run foroxalate and citrate were less than 1％ and 3.0％ and the betweenrun relative standard deviations were less than 2.0％ and 4.0％,respectively.The detection limits were 1 mg/L for both oxalate and citrate.The linearity ranges of oxalate and citrate were both 0-500 mg/L with the correlation coefficient between 0.999 5 and 0.995 4 (P ＜ 0.05),respectively.The average recoveries were 102.38％ for oxalate and 92.74％ for citrate.Conclusion This method is proved to be simple,sensitive and accurate,and also applied to determine oxalate and citrate in urine samples with satisfactory results.

Objective To investigate the effect of GDNF on pancreatic cancer cell proliferation and chemotaxis.Methods The cell counting, MTT and flow cytometry were employed to investigate whether the neurotrophic factor GDNF can stimulate the proliferation of pancreatic carcinoma cells.Meanwhile, the trans-well invasion chamber was used to observe the chemotactic effect of GDNF on pancreatic cancer cells.Results The cells were exposed to incremental concentrations of human re-combinant GDNF (0-120 ng/mL).We found that the proliferation of pancreatic cancer cells was stim-ulated by GDNF in a dose-dependent manner and the number of cells in "S" phenotype was increased;The count of cells was increased by GDNF in a dose-dependent manner.Conclusion GDNF can stimu-late the proliferation and invasion of pancreatic cancer cells in a dose-dependent manner.

Objective The truncated fibronectin-binding proteins A (Fba protein) were cloned and expressed, then animals were immunized with Fba protein and subsequently challenged with group A streptococcus (GAS) to further investigate protective antibodies induced by each domain and determine the most immunogenic domain. Methods Fba proteins, which were divided into four overlaps based on the structural domains, were truncated and expressed. The fba truncated genes were amplified by polymerase chain reaction (PCR) with SSI-9 of GAS as template, and cloned into prokaryotic expression plasmid pGEX-2T and expressed in E. coli BL-21. The products were confirmed by Western blot and purified by affinity chromatography column. Female BALB/c mice were immunized with the four truncated proteins respectively, with phosphate buffered solution (PBS) as control. The serum IgG of mice was detected by enzyme-linked immunosorbent assay (ELISA). After the third immunization, mice were challenged with fatal dose of GAS (M+ Fba+ ) to evaluate the protective rates in each group. The data were compared by analysis of variance and Fisher's exact test.Results Prokaryotic expression plasmids of pGEX-2T/FbaAl, pGEX-2T/FbaA2, pGEX-2T/FbaA3 and pGEX-2T/FbaA4 were successfully constructed and the four truncated proteins were expressed and purified successfully. Serum levels of IgG in each experimental group gradually increased with immunization with Fba protein more times. After the third immunization, the IgG titer against FbaA2 1290.2, P<0. 01). After GAS challenge, four out of eight mice were protected in FbaA2 protein group, while two out of eight mice in FbaAl protein, FbaA3 protein and FbaA4 protein groups,respectively (P<0. 05). Conclusions Four truncated Fba proteins are constructed and expressed successfully. Truncated FbaA2 protein could be able to induce strongest protective immune response.