Based on M9 culture medium,the concentration of ingredients of culture medium was optimized for the fermentation of pNSR32/BL21(DE3),the engineering bacterial with spider silk protein,and lactose as an inducer.The condition of optimum culture medium was obtained for the expression of the high molecular weight recombinant spider silk protein by using orthogonal and individual factor experimental design.The result was showed that the optimum culture medium was consisted of 0.3% glycerol,3% yeast,0.75% tryptone,0.05%(NH_4)_2SO_4 and a little inorganic salt_.It was confirmed that the optimum culture medium will benefit the growth of bacterial and expression of recombinant spider silk protein.The production level of propose protein will attain at 20% of the total proteins in the fermentation.

Objective To explore the effect of recombinant human vascular endostatin(Endostar,YH-16) on MCF-7 human breast cancer cells in nude mice. Methods Forty nude mice were randomized into four groups and received injection with MCF-7 cells. When gross tumor volume reached 1.0 cm3 ,mice were injected subcutaneously once every two days as following for 20 days: (1) O. 9% NS(0. 2 mg/kg) in negative control group;(2) YH-16(20 mg/kg) in YH-16 group; (3) DDP (1 mg/kg) in positive control group; (4) DDP (1 mg/kg) plus YH-16(20 mg/kg) in DDP plus YH-16 group. Mice were killed by cervical spine disloca-tion. And the body weight, tumor volume, rate of tumor inhibited, VEGF, apoptosis rate of tumor cells and pul-monary metastasis rate were detected. Results The means (SD) of tumor volume in DDP plus YH-16 group,positive control group, YH- 16 group and negative control group were 0. 686 (0. 229), 1. 258 (0. 101), 1. 888 (0. 215), 3. 366 (0. 284) cm3, respectively (P <0. 05) ; the rates of tumor inhibited in the four groups were 92. 1% ,57.3 %, 36. 5 % and 0, respectively (P < 0. 05) ; pulmonary metastasis rates were 0, 10%, 20% and 90% ,respectively(P <0. 05) ;the mean (SD) of apoptosis rates of tumor ceils were 31.6% (2. 7%) ,28. 1%(2. 7%), 19. 4% (2. 9%), 15.7% (3.2%), respectively (P < 0. 05). The mean of absorbance of VEGF in the four groups were 0. 530 ± 0. 164,0. 759 ± 0. 210,1. 063 ± 0. 295,1. 268 ±0. 145 (RT-PCR) and O. 260 ±0. 082,0. 348 ±0. 085,0. 461 ±0. 099,0. 556 ± 0. 113(Western blot) ,respectively(P < 0. 05). Conclusion YH-16 can inhibit MCF-7 breast cancer cell growth and distant metastasis in nude mice with little adverse reac-tion. YH-16 has synergistic effect combined with DDP.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by memory loss and cognitive decline. The pathological eatures of Alzheimer's disease are abnormal deposition of amyloid beta-peptides (Aβ), neurofibrillary tangles, cholinergic deficits, and loss of synaptic processes and dendritic spines. Senescence accelerated mouse prone 8 exhibits age-related deficits of learning and memory from an early age, tau protein phosphorylation, neurotransmitter changes, synaptic structure and function disorders, circadian rhythm disorders,as well as gene expression and many other characteristic changes, which are consistent with Alzheimer's disease pathological changes，and can be used as an ideal animal model for Alzheimer's disease prevention drugs development.

Objective To discuss the clinical effect of urinary kallikrein in the treatment of progressive cerebral infarction and its influence on hs -CRP.Methods 92 patients with progressive cerebral infarction were ran-domly divided into the observation group(n =46)and the control group(n =46)according to the digital table.The control group was treated with conventional therapy,the observation group was treated with urinary kallikrein based on the control group.The clinical effects and the influence on hs -CRP were compared between the two groups.Results The explicit efficiency and the effective rate of the observation group were 71.7% and 89.1%,which were significant-ly higher than 43.5% and 69.6% in the control group(χ2 =7.522 0,5.372 7,P <0.05 or P <0.01).After treat-ment,the hs -CRP levels in the two groups were (11.79 ±3.92)mmol/L and (10.04 ±3.90)mmol/L,which were significantly lower than (13.48 ±3.89)mmol/L and (13.54 ±3.93)mmol/L before the treatment(t =4.287 4, 2.075 5,P <0.05 or P <0.01),and hs -CRP of the observation group was significantly lower than the control group (t =2.146 5,P <0.05).After treatment,NIHSS scores in the two groups were (9.91 ±4.35)points and (6.82 ± 4.32)points,which were significantly lower than (15.39 ±4.34)points and (15.43 ±4.37)points before treatment (t =6.048 6,9.503 2,all P <0.01 ),and NIHSS score of the observation group was significantly better than the control group(t =3.418 5,P <0.01 ).After treatment,the Barthel in the two groups were (53.87 ±18.12)and (68.21 ±18.14),which were significantly higher than (34.35 ±18.08)and (34.42 ±18.11 )before treatment (t =5.172 1,8.943 4,all P <0.01),and Barthel of the observation group was significantly better than the control group(t =3.793 3,P <0.01).There was no obvious adverse reactions and side effects in the treatment.Conclusion Urinary kallikrein can effectively improve the local blood flow perfusion,restore nerve function damage and improve the living ability of the patients in the treatment of progressive cerebral infarction,it has curative effect,and it's safe and reliable,which is worthy of promotion.

A 27-peptide,a fragment of hepatitis delta antigen(HDAg),was synthesized and used to develop an ELISA method for ihe detection of anti-HD.It was found that positive anti-HD reaction occurred between the coated 27-peptide and a stored sample of serum which was known anti-HD positive.Absorption test revealed that the synthetic peptide competed with natural HDAg for anti-HD,suggesting that the peptide possessed the antigenicity similar to that of natural HDAg.The antigenicity of the synthetic peptide was quite specific wihtout cross reaction with normal human and mouse sera and with anti-HA.anti-HB and anti-HC sera.Among 300 blood donors,there was only 1 case(0.33%)anti-HD positive with an ALT level 2 times higher than normal.In 41 cases of non-B hepatitis and 52 cases of HAV hepatitis,none was anti-HD positive.In 211 cases of various types of HBV hepatitis,21 were(9.95%)anti-HD positive,among whom 2/82(2.5%)werehealthy HBV carriers,6/43(13.95%)were patients with a-cute icteric hepatitis,6/60(10.00%)were patients of chronic active hepatitis,4/18(22.20%)were patients of severe hepatitis,and 3/8(37.50%)were those with liver cirrhosis.These results were consistent with those in our previous reports.

Objective To evaluate the efficacy and safety of emergency percutaneous coronary intervention(PCI) in patients with non-ST-segment elevation acute coronary syndrome(ACS).Methods A total of 233 patients(emergency group) were treated with emergency PCI within 48 h of heart attack and another 152 patients(delayed group) were treated with PCI after 3-14 days of medical therapy.All culprit lesions were treated.Procedural success rate,the time from admission to angina relief,the length of hospital stay and cardiac events incidence in 30 days were observed.Results The procedural success rates for the emergency group and the delayed group were similar(98.1% vs 95.5%),but cardiac events incidence in 30 days was significantly lower in the emergency group than that of the delayed group(2.9% vs 14.1%,P

MicroRNAs (miRNAs) are evolutionarily conserved small noncoding RNAs of-22 nucleotides that exist in a wide variety of organisms,including animals,plants and virus.Mature miRNAs are able to control gene expression at a post-transcriptional level,either by blocking mRNA translation or inducing their degradation.Hepatitis B virus X protein (HBX) is a 17000 protein that is implicated to play a crucial role in hepatocarcinogenesis.Recently,many studies have shown that HBX is associated with miRNA regulation,and is involved in regulating fundamental biological processes of tumor in cell proliferation,differentiation and apoptosis.

Antiporters ; genetics ; Base Sequence ; Female ; Glycogen Storage Disease Type I ; diagnosis ; genetics ; Humans ; Infant ; Monosaccharide Transport Proteins ; genetics ; Mutation ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo observe the effect of Modified Yiqi Chutan Recipe (MYCR) on blood flow perfusion in treating mid-late stage non-small cell lung cancer (NSCLC) patients by using multislice CT perfusion (CTP) , and to assess the relationship between each CTP parameter and the prognosis as well.

METHODSTotally 87 mid-late stage NSCLC patients were randomly assigned to the treatment group (44 cases, Shenyi Capsule + MYCR +chemotherapy) and the control group (43 cases, chemotherapy alone) in the ratio of 1:1. And 21 days consisted of 1 therapeutic course, 4 courses in total. All of them underwent CTP of primary tumor and routine thoracic CT examination (plain CT and enhancement CT) 3 times (before therapy, after 2 and 4 cycles). CT findings were analyzed for tumor size and perfusion parameters [blood flow (BF), blood volume (BV), permeability surface (PS), mean transit time (MTT), and time to peak (TP) before and after treatment, and relationship between perfusion parameters and prognosis was also assessed.

RESULTSIn 87 cases, 7 dropped out and 80 cases were available, 40 in the treatment group and 40 in the control group. (1) The relief rate was 47.5% (19/40) and the total stable rate was 77.5% (31/40) in the treatment group, and they were 40.0% (16/40) and 65.0% (26/40) in the control group, with no statistical difference between the two groups (χ² = 0.672, 1.227; P > 0.05). (2) Compared with before treatment group in the same group, BF and PS decreased, and MTT increased in the two groups after 2 and 4 courses (P < 0.05); BE and PS decreased, and MTT increased in the control group after 2 courses (P < 0.05). Compared with the control group after 4 courses, BE decreased more significantly in the treatment group (P < 0.05). (3) After 4 courses, all patients were assigned to the remission group (35 cases) and the non-remission group (45 cases) according to the RECIST standard. Compared with before treatment in the same group, BF, BF, and PS all decreased, and MTT increased in the remission group after treatment (all P < 0.05); BF increased in the non-remission group after treatment (P < 0.05). (4) All patients were assigned to the BE increase group (34 cases) and the BE decrease group (46 cases) according to changed BE values after treatment. Results showed the mean survival rate was 246 days in the BF increase group (the 1-year accumulative survival rate being 13.0%) and 387 days in the BE decrease group (the 1-year accumulative survival rate being 53.1%). The life span was prolonged and the 1-year accumulative survival rate was elevated in the BE increase group, with statistical difference as compared with the BE decrease group (χ² = 19.057, P < 0.01).

CONCLUSIONSShenyi Capsule plus MYCR could reduce BE in mid-late stage NSCLC patients , improve vascular permeability, showing better synergistic effect with chemotherapy. CTP could not only reflect the change of tumor size, but also reflect vascular function of the tumor. Meanwhile, changes of CTP parameters were closely associated with prognosis. Patients with post-treatment decreased BE value had better prognosis and longer life span.

Capillary Permeability ; drug effects ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Phytotherapy ; Tomography, X-Ray Computed

Objective: To discuss the imaging features of esophageal fistula and the value of intervention.Methods: We retrospectively analyzed the complete clinical data of 12 cases of esophageal fistula treated from June 2000 to May 2008,of whom,2 developed esophago-thoracis fistula and esophago-mediastinum fistula respectively after conservative radiotherapy,and 10 developed esophageal fistula after surgery for esophageal cancer.All the patients received iodine-based oral contrast,spiral CT scanning and intraperitoneal stent implantation for fistula closure.Results: Iodine-based oral contrast showed that the contrast agent overflowed into the chest in 8 cases,into the airway in 3 and into the mediastinum in 1.Spiral CT scanning revealed 8 cases of pleural fistula and 2 cases of remnant stomach-airway fistula,but failed to display the other 2 because of the small size.Intraperitoneal stent implantation was successfully accomplished in all the cases,which helped to close the fistula,take in food,stop bucking,control lung,mediastinum and other infections,and improve the patients' quality of life.Conclusion: Esophageal fistula can be effectively diagnosed based on the findings of iodine-based oral contrast and spiral CT scanning.As a novel technique,intraperitoneal stent implantation for fistula closure,simple,safe and with obvious immediate effect,is well worth promoting.