Kallistatin (Kal) is a negative acute phase endogenous protein which can inhibit tumor angiogenesis, growth and metastasis effectively. To express and purify recombinant human kallistatin (rHKal), and characterize its biological activity, P. pastoris was transformed with pPIC9-Kal/GS115 (His4) to express rHKal. The fermentation was carried out in a 7.5 L bioreactor with high density cell culture. 1%-2% methanol was added to the medium to induce the expression of rHKal. The secretion was purified with phenyl sepharose, G-25 sepharose, heparin sepharose and Sephacryl S-100 chromatography. The biological activity of purified bulk rHKal on HUVEC was evaluated with MTT and tube formation assays. The final expression of rHKal in the supernatant reached 50 mg x L(-1), the purity of bulk rHKal after purification was above 98%. A dose-dependent inhibition of rHKal on HUVEC proliferation was observed, however, a U-shaped dose-response curve of rHKal on capillary formation of HUVEC was revealed. The described protocol provides an effective means for preparing rHKal that could be used for anti-angiogenesis therapy in the future.

Objective To explore the value of the spectral CT iodine-based material decomposition technique in differential diagnosis of central pulmonary carcinoma from obstructive pneumonia and atelectasis.Methods 25 cases with central pulmonary carcinoma complicating with obstructive pneumonia and atelectasis underwent CT plain scan and spectral contrast scan including pulmonary-arterial-phase (PAP) and bronchial-arterial-phase(BAP).Conventional CT images and iodine base images were generated in each phase by GSI viewer.The 4 groups of images differences between the tumor and the obstructive pneumonia and atelectasis were analyzed.Results The margin of the tumor was ill-defined in plain scan.The cases that showed difference between the tumor and the obstructive pneumonia and atelectasis in 4 groups of images were respectively as follows,10 in conventional CT images of PAP,1 6 in conventional CT images of BAP,1 9 in iodine base images of PAP,and 23 in iodine base images of BAP.The difference of the 4 groups was significant (χ2 =16.54,P<0.05).There was the highest accuracy in iodine base images of BAP in defining the tumor margin (χ2 = 12.83,χ2 = 4.20,χ2 =1.34,P <0.05). The accuracy between conventional CT images of BAP and iodine base images of PAP had no significant differences (χ2 =0.86,P >0.05),but were better than that of the conventional CT images of PAP(χ2 =6.65,P <0.05).Conclusion Spectral CT iodine-based material decomposition technique is helpful to improve subjective diagnosis of central pulmonary carcinoma complicating with obstructive pneumonia and atelectasis,especially in iodine base images of BAP,it can provide accurate information for clinical stage and treatment.

Objective To explore the main influence factors(heart rate,rotation speed,and reconstruction algorithm)on the image quality of coronary artery with 40 mm VCT(64-detector row helical CT)using a pulsating cardiac phantom.Methods An adjustable pulsating cardiac phantom(GE) containing predetermined simulated coronary arteries was scanned using a 40 mm VCT(GE LightSpeed CT) with cardiac pulsating rates of 40,45,50,55,60,65,70,75,80,85,90,95,100,105,110,and 115 beats per minute(bpm).The variable rotation speeds technique of 0.35 s,0.40 s,and 0.45 s were used, respectively.The raw data were reconstructed using both one-sector and multi-sector reconstruction algorithm at optimal window of the R-R interval.The image quality score(IQS)was evaluated by two radiologists according to the same evaluation standard of reformated image.The correlation between heart rate(HR), roation speed,reconstruction algorithm,and IQS were analyzed.The IQS as independent variable and the HR,rotation speed,reconstruction algorithm as dependent variables were analyzed by multiple linear regression analysis.Restllts The heart rate and the reconstruction algorithm had significant influence on IQS.The rotation speed(0.35s,0.40s,and 0.45s)didn't have significant influence on IQS.There was linear regression relationship between heart rate,reconstruction algorithm and IQS(P

Objective To systematically evaluate the efficacy and safety of nucleo(s)tide analogues (NAs)sequen-tial/NAs sequential combined with pegylated interferon (Peg-IFN)for the treatment of HBeAg-positive chronic hepatitis B(CHB).Methods PubMed,Cochrane Library,Embase,and Chinese Medical databases (CNKI,Wan-fang and VIP)from database establishment to March 25,2017 were retrieved,randomized controlled trials of NAs sequential/sequential combined with Peg-IFN for the treatment of CHB after application of NAs to achieve virologic response were included in study,Meta analysis was performed by RevMan 5.3 software,HBeAg seroconversion rate and HBsAg negative conversion rate at the end of treatment were compared.Results Nine studies were eventu-ally included,4 were about NAs sequential Peg-IFN,5 about NAs sequential combined with Peg-IFN.At the end of treatment,compared with using NAs monotherapy for antiviral treatment,NAs sequential/sequential combined with Peg-IFN therapy can improve HBeAg seroconversion rate(31.2% vs 11.7%;OR,3.69 [95%CI ,2.43 -5.60];P < 0.01 )and HBsAg negative conversion rate(11.5% vs 0.5%;OR,9.31 [95%CI ,2.72 - 31.89];P <0.01).According to the results of subgroup analysis,HBeAg seroconversion rate in NAs sequential Peg-IFN therapy group was higher than control group (25.3% [42/166]vs 10.0% [17/170];OR,3.1 [95%CI ,1.66 -5.79];P <0.01);HBeAg seroconversion rate in NAs sequential combined with Peg-IFN therapy was higher than control group (36.8%[63/171]vs 13.5%[23/171];OR,4.24[95%CI ,2.41 -7.46];P <0.01).Sequential/se-quential combination therapy showed more adverse reaction,most of which can be tolerated or improved after symp-tomatic treatment.Conclusion For the treatment of HBeAg-positive CHB,after application of NAs to achieve viro-logic response,NAs sequential/sequential combined with Peg-IFN therapy for 48 weeks can significantly increase HBeAg seroconversion rate and HBsAg negative conversion rate.

Kallistatin (Kal) is a negative acute phase endogenous protein which can inhibit tumor angiogenesis, growth and metastasis effectively. To express and purify recombinant human kallistatin (rHKal), and characterize its biological activity, P. pastoris was transformed with pPIC9-Kal/GS115 (His4) to express rHKal. The fermentation was carried out in a 7.5 L bioreactor with high density cell culture. 1%-2% methanol was added to the medium to induce the expression of rHKal. The secretion was purified with phenyl sepharose, G-25 sepharose, heparin sepharose and Sephacryl S-100 chromatography. The biological activity of purified bulk rHKal on HUVEC was evaluated with MTT and tube formation assays. The final expression of rHKal in the supernatant reached 50 mg x L(-1), the purity of bulk rHKal after purification was above 98%. A dose-dependent inhibition of rHKal on HUVEC proliferation was observed, however, a U-shaped dose-response curve of rHKal on capillary formation of HUVEC was revealed. The described protocol provides an effective means for preparing rHKal that could be used for anti-angiogenesis therapy in the future.
Bioreactors ; Capillaries ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Electrophoresis, Polyacrylamide Gel ; Fermentation ; Human Umbilical Vein Endothelial Cells ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Serpins ; biosynthesis ; genetics ; pharmacology

OBJECTIVETo analyze the differential proteomics in human umbilical cord mesenchymal stem cells (MSC) induced by chemical hypoxia-mimetic agent cobalt chloride (CoCl(2)) by two-dimensional gel electrophoresis (2-DE) and mass-spectrometry.

METHODS2-DE was performed to separate proteins from treated and untreated human umbilical cord MSC with CoCl(2). 2-DE images were analyzed by ImageMaster 2D Platinum software 6.0. The differential expressed proteins was identified by MALDI-TOF-MS. The differential proteins were classified based on their functions.

RESULTS2-DE reference patterns of CoCl(2) treated human umbilical cord MSC were established. A total of twenty-six differential proteins were identified, of them eleven proteins were up-regulated and fifteen down-regulated. Their biological functions involved in carbohydrate metabolism, protein metabolism and modification, lipid metabolism, coenzyme and prosthetic group metabolism, cell cycle, immunity and defense, cell structure and motility, signal transduction, protein targeting and localization, neuronal activities, muscle contraction, etc. Peroxiredoxin1 (Prdx) was down-regulated, whereas alpha-enolase (ENO1) and vesicle amine transport protein 1 homolog (VAT1) up-regulated.

CONCLUSIONThe effects of hypoxia on human umbilical cord MSC were participated by multiple proteins and involved in multiple functional pathways.

Cobalt ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; Proteome ; analysis ; Proteomics ; Umbilical Cord ; cytology ; drug effects

AIM:To investigate the mutation of FLT3-ITD gene in the patients with newly diagnosed acute myeloid leukemia (AML). METHODS:From March 1, 2015 to June 1, 2017, 207 patients with AML admitted to de-partment of hematology, sichuan provincial people′s hospital were enrolled in this study. The bone marrow samples were collected from the patients. PCR was used to detect the mutation of FLT3-ITD gene. After the corresponding chromosome was obtained by R-banding, the cells were made into strips and banding. Twenty karyotypes with relatively cleavage were automatically selected from each specimen to complete karyotyping. By analysis of the clinical data and following-up the prognosis, the FLT3-ITD gene mutation in diagnostic and evaluative values for AML were performed. RESULTS:FLT3-ITD gene mutation was found in 42 cases of 207 AML patients, the positive rate was 20. 29 % . FLT3-ITD positive patients showed 3 bands. FLT3-ITD gene mutation in 42 patients with positive results showed that FLT3-ITD gene mutations in turn met the end to end, and insert a number of nucleotides, but all the mutations were in-frame mutations. According to the FAB and WHO standard, in 42 cases of FLT3-ITD positive positive patients, M0 accounted for 0.00% , M1 accounted for 2.38% (1/42), M2 accounted for 23.81% (10/42), M3 accounted for 0.00% , M4 accounted for 2.38% (1/42), M5 accounted for 69.05% (29/42), M6 accounted for 0.00% , M7 accounted for 2.38% (1/42). The white blood cell ( WBC) level and complete response (CR) rate in FLT3-ITD positive patients were lower than those in FLT3-ITD negative patients (P<0.05). CONCLUSION:The WBC level and CR rate, which are lower in FLT3-ITD positive patients than those in negative patients, are the clinical risk factors. It will be helpful to determine the prognosis evaluation for AML pa-tients.

Objective To assess the curative effect of percutem transilluminated with negative pressured on the potaried technique on the treatment of venous ulcer in lower extremity.Methods The clinical date of 300 cases involving 300 legs with venous ulcer in lower extremity,who underwent the percutum transilluminated negative pressured potaried technique using TRIVEXTM Ⅱ potaried system or the percutum transfixion surgical treatment from October 2005 to June 2009,were analyzed.Three hundred cases were randomly divided into potaried group and transfixion group.In potaried group,there were 190 cases involving 190 legs treated with TRIVEXTM Ⅱ potaried system.In transfixion group,110 cases involving 110 legs treated with percutum transfixion.The clinical indexes of skin infection rate and skin necrosis rate,shrinkage rate of wound area and skin depigmentation rate,ulcer healing rate and ulcer recurrence rate were calculated to assess the clinical curative effect on day 5,day 20,day 120 and day 360 after operation respectively.Results The rates of skin infection and skin necrosis were significantly decreased in potaried group compared with transfixion group on day 5 after operation (P0.05).Ulcer recurrence rate was remarkably lower in potaried group than that in transfixion group on day 360 (P

Objective To study the epidemiologic characteristics of trauma in the prehospital first-aid in megapolis. Methods The epidemiologic data of 10 654 traumatic patients,including treated by prehospital treatment and emergency treatment from January 2000 to January 2005,were analyzed.Results The proportion of male was 70.96%,adult patients(21～50 years old)79.23%,suburb 62.86%,downtown 37.14%.The patients whose ISS scores surpassed 16 accounted for 37.98%,which caused by traffic accident was 37.74%,by public order 24.39%,by industrial trauma 21.71%.The trauma in the limbs accounted for 67.51%,cephalic and cervical wounds accounted for 58.64%,multiple wounds 41.77%,thoracic and abdominal wounds 39.41%.Three hundred and sixty one were killed on the spot,which caused by traffic accident were 46.81%,by public order 28.81%,by industrial trauma 14.40%.Forty-two percent point six six patients died of multiple trauma,54.07% died of cephalic and cervical trauma,15.79% died of thoracic and abdominal wounds.Conclusion Suburban area gradually became the frequently-occurred areas of trauma in megapolis. The wounded were mainly young adults and had a tendency of juvenility.The majority of damaging and lethal factors were traffic accident,public order and industrial trauma.Some pertinet measures and professional first-aid models may improve the traumatic first-aid level.

Abstract: Objective　To master the condition of cockroach population distribution, seasonal dynamics, cockroach density for different habitat, and to provide a basis for developing cockroach control strategies. Methods　Six types of surveillance sites, including residential areas, hotels, restaurants, supermarkets, hospitals and farm product markets, were set up in 14 cities in Liaoning to monitor cockroaches using the sticky-trap method. The cockroach surveillance data from vector surveillance sites in fourteen cities of Liaoning Province in 2021 were collected and statistically analyzed using Excel 2010 and SPSS 23.0 software. The density and species composition of cockroaches were analyzed, and the density difference and seasonal dynamics trend of cockroaches in different habitats were compared. Results　A total of 3 031 cockroaches were captured in 2021, of which Blattella germanica accounted for 94.66% (2 869/3 031) and was the dominant population. The total density of cockroaches was 0.230 0 cockroaches per sheet (3 031/13 234) and the total infestation rate of cockroaches was 5.59% (562/10 052). The density and infestation rate of cockroaches in different habitats were in the order of farm product markets, restaurants, and hotels and the difference in infestation rate between habitats was statistically significant (χ2=168.327, P<0.05). The seasonal dynamics trend of cockroach density and disoperation rate showed a unimodal curve, and the peaks were distributed in July. The seasonal dynamics of cockroach density and disoperation rate in different habitats showed a unimodal curve in the habitats of farm product markets, supermarkets, hotels, hospitals and residential areas all, while the habitats of restaurants were close to a double peak curve. Conclusions　B. germanica is the dominant species of cockroaches in Liaoning Province in 2021. Compared with 2020 the density and disoperation rate of cockroach in 2021 showed a slight downward trend, and the seasonal dynamics trend of cockroach density and disoperation rate showed a unimodal curve. The farm product markets are the key places for cockroach prevention and control. According to the seasonal fluctuation trends in cockroach density and infestation rate in different habitats, comprehensive prevention and control measures should be taken before the peak periods to reduce cockroach density and control diseases.