The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.

Objective To observe the clinical efficacy of acupoint application withChan Wu gel cream plus five notes therapy in treating cancer pain.Method Eighty patients with cancer pain were randomized into a treatment group and a control group, 40 cases in each group. The treatment group received acupoint application withChanWu gel cream at the corresponding acupoints of the affected organ in association with the five notes therapy; the control group was intervened by the three-step analgesic ladder recommended by World Health Organization (WHO). The quality of life and pain intensity were scored before and after the intervention.Result The quality of life and pain degree were significantly changed after the treatment in the two groups (P<0.01). The quality of life and pain intensity of the treatment group were significantly different from that of the control group (P<0.05).Conclusion Acupoint application withChan Wu gel cream plus five notes therapy is effective in mitigating cancer pain and improving the quality of life.

Background and purpose:Ara-C is one of the most effective and common agents in the treatment of acute nonlyphocytic leukemia. Telomerase is a unique complex of ribonucleoprotein. It plays an important role in the pathogenesis and development of cancer. In this study, we investigate the changes of mRNA expression of telomerase subunits in HL-60 cells induced by Ara-c and try to come up with a theory that could help to assess the efficacy of Ara-C. Methods:The combinations of various Ara-C concentration and the incubation time were used to treat HL-60. The ratios of apoptotic cell to necrosis cell were determined by flow cytometry and the expressions of telomerase subunits mRNA were evaluated by RT-PCR.Results:① There was no influence on transcription of telomerase subunits gene after HL-60 cells was cultured with 0~0.2ug/ml Ara-C for 12 hours;② 2ug/ml and 10ug/ml of Ara-C could down regulate the expression of hTERT from 0.80+0.07 to 0.50+0.04 and 0.39+0.03, not hTR and hTP1;③ with longer incubation with 10ug/ml of Ara-C, the percentage of apoptosis could be increased. The maximal induction of apoptosis (18.16+4.25%) could be reached at 12hrs treatment of Ara-C, then gradually decreased later on. The rate of necrosis increased with time, the maximal percentage(57.94+12.03%) of necrosis was observed at 48hrs of incubation time with drug. The mRNA level of hTERT gene also decreased along with the cultured time , the lowest value (0.18+0.03) has been documented at 48hrs time point, but not hTR、TP1.Conclusions:① Ara-C could down-regulate the expression of hTERT mRNA in a dose-and time-dependent manner, but not hTR、hTP1;② There might be no relationship between the percentage of apoptosis induced by Ara-C apoptosis and the expression of telomerase hTERT gene mRNA, but a close relationship between necrosis and the expression of hTERT mRNA has been found.

Objective The purpose of this study was to investigate the clinical significance of serum vascular endothelial growth factor(S-VEGF)in patients of esophageal squamous cell carcinoma with the end-point of progress-free survival rate.Methods Sera from 89 patients with esophageal squamous carcinoma,who presented and treated with concurrent chemoradiotherapy or surgical resection in Cancer Center d Sun Yat-Sen University from December 2002 to May 2004,were sampled at the time of pre-treatment.30 cases of health individuals without any evidence of disease were selected as control group.For patients with surgical resection were performed with esophagectomy and extended lymphadenectomy.For patients with concurrent chemoradiotherapy comprised of cisplatin and 5-fluorouracil.The radiotherapy dose of 2 Gy per day was initiated on Day 1 of chemotherapy and continued daily for 5 days per week for 6 weeks,for a total close of 60 Gy.Two courses of chemotherapy were given during radiotherapy at 6-week intervals.The serum vascular endothelial growth factor(S- VEGF)levels were measured with a solid phase enzyme Human VEGF Immunoassay ELISA kit.The end point of this study was progress-free survival,and time was calculated from the date of diagnosis to date of relapse or last follow evaluation.Results S-VEGF level of patients with esophageal squamous cell carcinoma was significantly higher than that of heahhy controls(475.93?44.76 pg/ml vs.294.20?23.40 pg/ml;P=0.020).S-VEGF levels in patients with StageⅢand StageⅣdisease were significantly higher than those in patients with Stage I and StageⅡdisease.The 1-year progress-free survival rate d high S-VEGF group(≥475 pg/ml)was significantly lower than that of the low S-VEGF group(＜475 pg/ml)(24% vs.66%;P=0.0004).The 1-year progress-free survival rate of the patients with StageⅠand StageⅡdisease was significantly higher than that of patients with StageⅢand StageⅣdisease(71% vs.29%;P=0.0015).The variables were assessed by multivariate analysis using the Cox proportional hazards model,and the results revealed that the clinical stage and the S-VEGF were first and second independent prognosis factor,respectively.Conclusions In the current study,a high S-VEGF was found to be associated with tumor progression,poor treatment response,and poor survival in patients with squamous cell carcinoma of the esophagus.

Objective To evaluate the significance of CT assessment for extraarticular anatomy in treatment of displaced intraarticular calcaneal fractures.Methods(1)Measurement of normal calcaneum 40 pieces of adult calcaneum specimen were measured,items of measurement included height of culmination of posterior facet and tuberosity,width of posterior edge of sustentaculum and tuberosity.(2)CT measurement of calcaneum.Transverse(axial)and coronal CT scanning were obtained from 20 feet with displaced intraarticular calcaneal and 20 normal feet as control.Following items were measured in CT scanning:the height of culmination of posterior facet and tuberosity,the coronal talocalcaneal angle,in coronal scanning,the width of posterior edge of sustentaculum and tuberosity,the axial calcaneocuboid angle,in axial scanning.Results(1)Measurement of height of calcaneum height of culmination of posterior facet and tuberosity of calcaneal specimen were(43.07?2.85)mm and(44.69?3.67)mm respectively,and these two items from CT scanning of normal feet were(42.84?1.66)mm,(43.40? 3.01)nun,and from CT scanning of feet with calcaneal fractures were(34.76?3.24)mm,(40.41? 3.69)mm.There was a statistically significant different between these two items for normal calcaneal specimen and for CT scanning of feet with ealcaneal fractures(P

Objective To find noninvasive estimation of partial pressure of carbon dioxide in artery (PaCO2) by measuring the end-tidal CO2 partial pressure (PETCO2) in elderly patients with respiratory failure with chronic obstructive pulmonary disease (COPD). Methods All the 30 acutely exacerbated COPD subjects received routine clinical treatment including bronchodilators,mucolytics, glucocorticosteroid, antibiotics and oxygen therapy for 5-7 days, and part of them received noninvasive positive-pressure ventilation (NIPPV) treatment concurrently. They were both tested by eupnea method and prolonged expiratory method before and after treatment. Results Before treatment, PET CO2 (Q) (end-tidal CO2 pressure with eupnea, (50.72±8.93) mm Hg wassignificantly lower than PaCO2 (F=38.73, P＜0.01 ). Yet, PETCO2(P) (end-tidal CO2 pressure with prolong expiration) was (70.35±8.91) mm Hg and PaCO2 was (71.25±9.08) mm Hg. There was no significant difference between PETCO2 (P) and PaCO2 (P＞0.05). The similar results were found after treatment. By linear regression analysis, PetCO2(P) was remarkably positive correlated with PaCO2 before and after treatment (r=0.96 and 0.97, respectively, P＜0.01). According to TABG,PCO2(C) which was calculated by the average expiratory time of fitting curve was (71.78±9.04)mm Hg. And there was no significant difference between PCO2 (C) and PaCO2 (P＞0.05);Thesimilar results were founcl after treatment. By linear regression analysis, PCO2 (C) was remarkably positively correlated with PaCO2 (r=0.97 and 0.98, respectively, P＜0.01) before and after treatment. Conclusions In COPD patients with type Ⅱ respiratory failure, conventional PETCO2 (Q) is significantly lower than PaCO2. Yet, PETCO2 (P) could exactly estimate PaCO2 and is appropriate to its dynamic monitoring. And PCO2 (C) which is calculated by mode Boltzmann on eupnea curve also obtain similar results.

OBJECTIVETo explore the association of 8-hydroxyguanine glycosidase OGG1 Ser326Cys polymorphism with semen quality and the risk of male infertility.

METHODSThis case-control study included 620 idiopathic infertile patients and 385 normal fertile controls. We determined their genotypes by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and analyzed their semen quality by computer-aided semen analysis (CASA).

RESULTSThe individuals with OGG1 326 Cys/Cys showed significantly lower sperm motility and concentration ([52.1 +/- 26.7]% and (3.75 +/- 0.91) x 10(6)/ml, ln transformed value) than the Ser/Ser carriers ([59.0 +/- 21.8] % and (4.12 +/- 0.88) x 10(6)/ml, ln transformed value) (P < 0.05). The risk of male infertility increased 69% in the OGG1 326Cys allele carriers as compared with the Ser carriers (OR = 1.69, 95% CI: 1.24 -2.31).

CONCLUSIONOGG1 326 Ser/Cys polymorphism might contribute to the risk of male infertility in the southern Chinese population.

Adult ; Case-Control Studies ; DNA Glycosylases ; genetics ; Genotype ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Semen Analysis ; Young Adult

Objective To investigate the effects of Helicobacter priori (H.prlori) strains from the patient with gastric cancer on the expression of DNA mismatch repair (MMR) genes in AGS cells in vitro. Methods AGS cells were co-cultured with ten strains of H.prlori from five patients with gastric cancer and five patients with gastritis respectively.The expressions of DNA MMR genes (hMSH2 and hMLH1) in mRNA and protein levels were determined by RT-PCR and Western blot.The enteropathogenic E.coli served as a bacterial control.Results E.coli and H.priori strains from gastritis have no effects on the expression of hMSH2 and hMLH1 in both protein and mRNA levels on the whole,while all the H.prlori strains from gastric cancer reduced expression levels of hMSH2 and hMLH1.Conclusions There are different effects between H. prlori strains from gastric cancer and those from gastritis on DNA MMR in AGS cell line,indicating that infec- tion of some H.prlori strains might lead to the inhibition of DNA MMR,and that in turns increases the risk of gastric carcinogenesis during chronic H.prlori infection.

Increased recognition of human parvovirus B19,as a significant human pathogen has resulted in intensive researches to understand the pathogenesis of B19 infection,to elucidate the nature of Th1-mediated cellular immune response,to improve diagnostic strategy that is deployed to detect B19 infection and blood-product contamination,and to lay a foundation that should contribute to the development of an effective vaccine to prevent B19 infection.In this review,the biologic characteristics and the pathogenesis of human parvovirus B19,and B19-related manifestations as well as laboratory diagnostic methods for B19 infection were comprehensively discussed.

The high expression of tissue factor (TF) is related to the coagulation disorder in acute leukemia. TF in blood circulation is mainly expressed in cells, microparticles (MP) and alternatively spliced human tissue factor (asHTF). To elucidate the role of TF in the coagulation disorder of acute myeloid leukemia (AML), RT-PCR was performed on 6 common AML cell lines NB4, HL-60, Kasumi-1, U937, K562 and THP-1. The results showed that only NB4 and U937 cells expressed baseline full-length TF and asHTF which were proved by sequencing. The flow cytometric detection, TF activity and TF antigen tests in NB4 and U937 cells revealed that the asHTF was expressed in trace amount and almost had no activity, while the TF antigen and activity in microparticles were significantly higher than that in asHTF. It is concluded that asHTF may play an unimportant role in the coagulation disorder of AML. Microparticle associated tissue factor (MP-TF) is the predominant source of TF activity released from AML cells.
Alternative Splicing ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; Thromboplastin ; genetics ; metabolism ; Tumor Cells, Cultured ; U937 Cells