To explore the clinical efficacy of Herba Saxifragae cream (HS), a compound of traditional Chinese herbal medicine, on chronic eczema.

To construct, express and purify fusion protein containing HBcAg and HBsAg preS1 epitope peptide for the purpose of investigating a novel HBV vaccine with both prophylactic and therapeutic functions. Using DNA recombinant technology, prokaryotic expression plasmid pBTcs1 expressing HBcAg and HBsAg pre-S1 epitope peptide fusion protein was constructed. After expressed in E.coli. HB101, the production BTcs1 was purified by sucrose density gradient ultracentrifugation and identified by SDS-PAGE, SEC, Western-blot and electron microscope. The results indicated that expression plasmid pBTcs1 was constructed successfully, and 20~25 mg purified BTcs1 fusion protein was obtained from 1L LB culture. Result of DOT-BLOT indicated that the distribution of BTcs1 was mainly in 30~50% sucrose, the purity of BTcs1 was greater than 95% by SDS-PAGE and SEC analysis. BTcs1 could probe with specific antibodies at 28 kDa by Western-blot, BTcs1 could also self assemble VLP by electron microscope analysis, its diameter was 30~34 nm approximately. The present study lay a foundation for further research functions and applications of BTcs1.

Nowadays,small peptides are always expressed in the form of fusion protein.The expression product contains many superfluous amino acids which can affect the biological functions of small peptides even expressed by GST fusion protein expression system.SUMO protease can cut SUMO fusion protein expressed by fusion expression system without any amino acid residues left on target protein thus become a hot topic in this field.Recombinant His-UlP1/pET3c/BL21(DE3)engineering strain was constructed by genetic engineering technology and the expression conditions were optimized in shake flaks.The process of high density fermentation was explored and different purification conditions were detected by chromatography.The results showed that SUMO protease could be expressed well after inducing the engineering strain by IPTG of 1.0mmol/L at 30℃ for 6 hours.The expression level of the strain in fermentation pots could reach 24.3% analyzed by SDS-PAGE.The purity of SUMO protease was more than 98% after further purification by cation exchange chromatography.The yield was 355mg SUMO protease per liter fermentation liquid.Western blot analysis demonstrated that there were immune reactions between IlP1 and 6?His antibodies,so it has established a good foundation for large-scale industralazation in the future.

Macrophages are involved in many important biological processes and membrane proteins are the key effector molecules for their functions. However, membrane proteins are difficult to analyze by 2-DE based method because of their intrinsic tendency to self-aggregate during the first dimension separation (IEF). To circumvent the obstacle hampering membrane protein analysis, we combined one-dimensional SDS-PAGE with capillary liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using this technique, we identified 458 GO annotated membrane proteins with extremely high confidence, including most known markers of peritoneal macrophages (e.g., CD11b, F4/80, CD14, CD18, CD86, CD44, CD16 and Toll-like receptor). Thirteen other CD antigens and 18 Ras-related small GTPase were also identified. In addition to those known macrophage membrane proteins, a significant number of novel proteins have also been identified. This research provides a valuable data set of macrophage membrane proteins, thus allowing for more comprehensive study of membrane proteins and a better understanding of the function mechanisms of macrophages in many biological processes.

OBJECTIVETo observe blood uric acid levels and Goldstein grading, as well as their correlation in Wilson's disease (WD) patients with different Chinese medical syndrome types.

METHODSTotally 906 WD patients in line with inclusive criteria were assigned to 6 groups, i.e., the heart spirit confused by phlegm group (HSCP, 26 cases), the phlegm-fire disturbing heart group (PFDH, 90 cases), the retention of damp-heat group (RDH, 113 cases), deficiency of qi and blood group (DQB, 168 cases), the deficiency of Gan-yin and Shen-yin group (DGYSY, 327 cases), the deficiency of Gan and Shen group (DGS, 182 cases) due to different Chinese medical syndrome types. Recruited were another 160 healthy subjects having similar ages and diet structures, who came for medical examinations, as the healthy control group. Venous blood was collected from the medial cubital vein of each-patient on an empty stomach in early mornings to detect blood uric acid levels. Results Blood uric acid levels were lower in each syndrome type group than in the healthy control group (146.08 +/- 67.24 micromol/L in the HSCP group; 157.08 +/- 69.77 micromol/L in the PFDH group; 162.58 +/- 97.72 micromol/L in the RDH group; 156.20 +/- 62.63 micromol/L in the DQB group; 161.83 +/- 111.23 micromol/L in the DGYSY group; 194.41 +/- 90.01 micromol/L in the DGS group; 242.39 +/- 87.55 micromol/L in the healthy control group, P < 0.01). Blood uric acid levels were higher in the DGYSY group than in the other 5 syndrome groups (P < 0.01). Correlation analyses between Goldstein grading and blood uric acid showed that, along with increased Goldstein grade (that was aggravating disease conditions), WD patients' blood uric acid levels decreased (P < 0.01).

CONCLUSIONSWD patient's blood uric acid levels decreased more. Blood uric acid levels and Goldstein grading were different in various Chinese medical syndrome types. Blood uric acid levels had certain value in assessing the severity of WD.

Asian Continental Ancestry Group ; Heart ; Hepatolenticular Degeneration ; blood ; classification ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Syndrome ; Uric Acid ; blood

Objective To investigate the effect of allogeneic transfusion and preoperative autologous blood donation on perioperative cellular immunity and humoral immunity in patients with gastrointestinal neoplasms.Methods Sixty gastrointestinal cancer patients undergoing radical surgery,33 males and 27 females,aged 53 69 years,weighing 47-70 kg,ASA physical status Ⅰ or Ⅱ,were included in this study.Blood transfusion was started when the blood loss was more than 200～400 ml,Hb＜70 g/L,and the patients were randomly divided into the preoperative autologous blood donation group (group P,n =30):intraoperative blood transfusion using the stored autologous blood transfusion;allogeneic blood transfusion group (group A,n =30):allogeneic blood transfusion.The levels of T lymphocyte subsets,NK cells and IL-2,IL-10,TNF-α,perforin (PF) concentration and plasma immunoglobulin IgG,IgA and IgM levels.Results The percentage of CD3+,CD4+,NK cells and the ratio of CD4+/CD8+ in group A at the end of surgery to 7 d after operation were significantly lower than those at the time of admission (P＜0.05).The percentage of CD3+,CD4+ (P＜ 0.05).The level of IL-2 in group A was significantly lower than that in group P (P＜0.05) 1-7 d after operation,and the level of IL-10 in group A was significantly higher than that in group P (P＜ 0.05).The levels of IgG and IgA 3 d after operation in group A were significantly lower than those in group P (P＜0.05).The levels of IgG and IgA in group P were significantly decreased at the end of operation and recovered to preoperative levels 1-3 d after operation (P ＜ 0.05).Conclusion Allogeneic blood transfusion can reduce the percentage of T-cell subsets and NK cells in cancer patients and delay their recovery,and also can transiently reduce the content of immunoglobulin IgG and IgA in plasma and thus affect the immune function of patients.However,the preoperative autologous blood donation has a slight effect on postoperative immune function in cancer patients.

OBJECTIVETo evaluate hemodynamic character and surgical effect of stenoses or occlusion of carotid artery system by perfusion-weighted MRI.

METHODSTwenty-one patients with stenoses or occlusion of carotid artery system underwent surgical treatment. Two patients underwent endarterectomy, extracranial-intracranial arterial bypass to 10 patients, and percutaneous stent angioplasty to 9 patients.

RESULTSPreoperative perfusion-weighted magnetic resonance (MR) revealed normal regional cerebral blood flow in all patients, with delayed time to peak in both middle cerebral artery distribution and border zone. Postoperative perfusion-weighted MR revealed normal time to peak in border zone, but abnormal in middle cerebral artery distribution. Evaluated by perfusion-weighted MR, it showed that surgical method can improve the hemodynamic disorder of this kind of disease. The early curative effect of endarterectomy and percutaneous stent angioplasty is better than extracranial-intracranial arterial bypass.

CONCLUSIONSPerfusion-weighted MR is a good method to evaluate hemodynamic character of stenoses or occlusion of carotid artery system. Surgical method is helpful to this kind of disease, and its curative effect can be evaluated impersonally and accurately by this new technique.

Adult ; Aged ; Angioplasty ; Carotid Stenosis ; physiopathology ; surgery ; Cerebrovascular Circulation ; Endarterectomy, Carotid ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Angiography ; methods ; Male ; Middle Aged ; Sensitivity and Specificity ; Treatment Outcome

OBJECTIVETo analyze the complications and treatment results of intraoperative radiotherapy (IORT) for esophageal carcinoma.

METHODSSixty patients with thoracic esophageal carcinoma underwent esophagectomy through right thoractomy, 30 patients of whom received IORT of 15 - 25 Gy.

RESULTSIn patients who underwent IORT, 2 cases of pneumonitis, 1 case of anastomotic leak and 1 case of incisional wound infection were found. In patients underwent surgery only, 1 case of thoracic empyema and 1 case of anastomotic leak were found. All the complications ultimately healed. There was no operative mortality. During the follow-up of 3 years, in patients who underwent IORT, 2 of 3 died of radiation pneumonitis 24 and 26 months after IORT with one complicated with bronchoesophageal fistula. One of 3 died of multiple lung metastases. The 3-year survival rate was 88.0% (22/25) in IORT group and 76.0% (19/25) in surgery only group.

CONCLUSIONIntraoperative radiotherapy can reduce locoregional recurrence if performed to thoracic esophageal carcinoma patients without surgical contraindication or distant metastasis. Radiation pneumonitis, a common complication difficult to manage, implies a poor prognosis and, consequently, the lung and bronchus should be protected from the radiation.

Adult ; Aged ; Combined Modality Therapy ; Esophageal Neoplasms ; mortality ; therapy ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Radiotherapy Dosage

BACKGROUNDCarotid artery stenting (CAS) as a competing treatment modality has had to adhere to limits to gain widespread acceptance in some studies. This study analyzed the clinical data of 1700 consecutive patients after CAS to retrospectively evaluate the 30-day outcome of CAS for internal carotid artery stenosis in a Chinese population.

METHODSMedical records of 1700 patients who underwent CAS at Xuanwu Hospital affiliated to Capital Medical University between January 2001 and August 2012 were reviewed. Postoperative 30-day complication rates were analyzed and compared with those of other studies. Univariate and multivariate Logistic regression analyses were used to identify factors associated with perioperation myocardial infarction (MI), stroke, and death.

RESULTSThe overall 30-day rate of MI, stroke, and death after CAS was 2.53%. In univariate analysis, patients who were symptomatic, had a neurological deficit (modified Rankin score (mRS) ≥3; P = 0.001), and who were not taking statins experienced a significantly increased rate of MI, stroke, and death (P = 0.017). In multivariate Logistic regression analysis, the presence of symptoms (odds ratio (OR) = 2.485; 95% confidence interval (CI) = 1.267-4.876; P = 0.008) and a neurological deficit (mRS ≥3) (OR = 3.025; 95% CI = 1.353-6.763; P = 0.007) were independent risk factors for perioperative MI, stroke, and death.

CONCLUSIONSAccording to this single-center experience, CAS may effectively prevent and treat carotid artery stenosis that would otherwise lead to stroke. Being symptomatic and having a neurological deficit (mRS ≥3) increased the risk of perioperative MI, stroke, and death.

Adult ; Aged ; Aged, 80 and over ; Carotid Stenosis ; surgery ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Myocardial Infarction ; Stents ; Stroke ; surgery ; Treatment Outcome

This study was aimed to investigate the expression and role of 14-3-3ζ in the AML cell lines: sensitive HL-60 and drug-resistant HL-60/VCR cells. Semi-quantitative RT-PCR and Western blot were respectively used to examine the expression of mdr1 mRNA and Pgp in AML cell lines to validate the results of microarray. Western blot was performed to investigate the expression of Pgp, 14-3-3ζ, and anti-apoptosis protein BCL-2, MCL-1 proteins. Immunofluorescence assay was used to detect the subcellular location of 14-3-3ζ protein in HL-60 and HL-60/VCR cells by laser scanning confocal microscopy. Transduction with siRNA was used to silence 14-3-3ζ in AML cell lines. Cell count method and flow cytometry of cell cycle were used to analyze the changes of growth of AML cells. The results found that mdr1 mRNA and Pgp did not expressed in HL-60 cells, but significantly overexpressed in HL-60/VCR cells. Except 14-3-3σ, the expression of other subtypes of 14-3-3 was higher in HL-60/VCR cells than that in HL-60 cells, especially 14-3-3ζ. The higher expression of 14-3-3ζ, BCL-2, MCL-1 protein was observed in HL-60/VCR cells than that in HL-60 cells. These results were same results from gene chip. It was also noticed that 14-3-3ζ was located in the cytoplasma and nuclei of AML cell lines, especially over-expressed in HL-60/VCR cells. Furthermore, suppression of 14-3-3ζ by RNA interference resulted in inhibition of the proliferation of AML cells with decreased protein expression of BCL-2 and MCL-1, especially in HL-60/VCR cells. It is concluded that 14-3-3ζ plays an important role in proliferation of AML cells and associates with BCL-2 and MCL-1 expression. These results suggested that development of therapy targeting 14-3-3ζ may provide novel, effective strategies for refractory and relapsed AML.
14-3-3 Proteins ; metabolism ; ATP Binding Cassette Transporter, Sub-Family B ; metabolism ; Apoptosis ; Cell Proliferation ; HL-60 Cells ; metabolism ; Humans ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism