The technique of double-layered plate was developed for screening the library of mutant endoglucanase III from Trichoderma reesei generated by the method of directed evolution.The enzyme activity was determined according to the velocity of the formation of halos on the plates.Several mutants with higher activity than the wild type at low temperature or alkaline pH were obtained by using this strategy under different screening conditions.Further results of spectrophotometric determination of the activities of these mutants were consistent with the results of plate screening.The establishment of such strategy will broaden the applications of the directed evolution methods for improving the existing proteins to obtain useful enzymes with new properties for industrial applications.

OBJECTIVETo characterize the immune reaction in SD rats exposed to trichloroethylene (TCE) and to identify the gene expression profiles involved in skin after TCE exposure.

METHODSFifteen percent of TCE was injected intradermally into the rat back (100 microL/120 g) at intervals of 7 days. Whole blood was collected 24 h after the fifth or seventh intradermic administration of TCE. The percentages of CD4+ and CD8+ of T lymphocytes were measured by a flow cytometer. The concentrations of IFN-gamma and IL-4 in the serum were semi-quantified by ELISA. Total RNAs of skin samples at 3 h or 24 h after the seventh dose of TCE in SD rats were extracted, and gene expression profiles of these tissues were analyszed by rat toxicology U34 array of Affymetrix.

RESULTSObvious decline of CD4+ in T lymphocytes was observed in the TCE-administer group. No significant concentration differences in IFN-gamma and IL-4 were found between TCE-treated and control rats. Gadd45a and Mel were significantly up regulated in skin tissue 24 h after TCE exposure. The expression regulation of immune response factors was as active as proteins associated with lipid metabolism and synthesis process in these skin samples of SD rats exposed to TCE.

CONCLUSIONT-helper type 1 cells mediate immune response can not be elicited in TCE-treated SD rats, but certain immune disorder can be induced.

Animals ; Female ; Gene Expression ; immunology ; Gene Expression Profiling ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; immunology ; Trichloroethylene ; immunology

Objective To evaluate the efficacy and adverse effects of gefitinib in the treatment of fe- male patients with advanced adenocarcinoma of lung who had failed to previous chemotherapy.Methods These patients received 250mg of gefitinib orally,once daily until disease progression or development of intol- erable toxic reaction.They were evaluated one month after treatment and every other month thereafter.Results Among the 27 evaluable patients,there were 1 CR(3.7%),11 PR(40.8%),10 SD(37.0%)and 5 PD(18.5%). The overall response rate was 44.5%(95% CI 29%～68%);and 22 patients(81.5%)gained profit(CR+PR+ SD)from the clinical therapy(95% CI 62%～94%);the mean TTP was 7.2 months.Symptomatic improvement rate was 80.0%.The main adverse effects were mild rash and diarrhea.Conclusion gefitinib has significant efficacy in the treatment of female patients with advanced tung cancer who had failed to previous chemother- apy.Adverse effects are mild.gefitinib is a suitable therapy for these patients.

Qigui Tongfeng tablet (QLTFT) is a traditional Chinese medicine with good effect for treating gout. Here, network pharmacology method and molecular similarity analysis were utilized to study the effective substance basis and molecular mechanism of the QLTFT on the gout. The similarity to the medicinal compounds is reflected in the Tanimoto coefficient that gives the structural similarity of two compounds. Operationally, similar modifiers were described as pairs of concepts with a similarity score of 0. 500. The results of the molecular similarity analysis suggested that the flavonoids in QLTFT could be new leads for gout. Furthermore, complex biological systems may be represented and analyzed as computable networks. Two important properties of a network were degree and betweenness. Nodes with high degree or high betweenness may play important roles in the overall composition of a network. And the results of network analysis showed that dongbeinine, verticinone-N-oxide, verticine N-oxide, peimine, peiminine, isobaimonidine, dongbeirine, peimisine and simi-arenol which with high degree acted on xanthine dehydrogenase/oxidase, matrix metalloproteinase-9, an arachidonate 5-lipoxygenase-activating protein, tyrosine-protein kinase and etc. Inhibition of these targets can prevent the formation of uric acid, reduce inflammation by uric acid and regulate the body's immune response. Thus, these compounds may be the main effective substance basis. The research results not only reveals its molecular mechanism, but also provide a theoretical basis for the quality control of drugs and clinical application.
Gout ; drug therapy ; Humans ; Medicine, Chinese Traditional ; Pharmacology ; methods ; Tablets ; Technology, Pharmaceutical ; methods

Adult ; Biopsy ; DNA ; genetics ; Female ; Gene Expression Profiling ; Glomerulonephritis, IGA ; genetics ; pathology ; Humans ; Kidney ; pathology ; Male ; Middle Aged ; Young Adult

Objective To investigate the significance of the serum levels of interleukin-10 (IL-10),IL-13,IL-15 of patients with chronic hepatic failure and the correlation between those inter- leukin levels and nosocomial infections.Methods The serum levels of IL-10,IL-13,IL-15 of 58 patients with chronic hepatic failure were measured by double antibody sandwich enzyme-linked immu- nosorbent assay at the time of admission and 2 weeks after admission.Results The serum levels of IL-15 and the propotion of IL-15/IL-10 and IL-15/IL-13 in patients with chronic hepatic failure group at the time of admission were significantly higher than those in healthy control group[(358.16?290.91) ng/L vs (38.55?21.49) ng/L,12.93?14.26 vs 1.10?0.55,98.55?97.5.5 vs 9.70?5.03,respectively,all P=0.000].Those in death group were significantly higher than those in improving group[(479.93v205.52) ng/L vs (244.51?236.29) ng/L,17.65?17.78 vs 8.53?7.98,130.69?115.50 vs 68.55?65.99,respectively,all P

OBJECTIVETo prepare and characterize the heavy metal reference material of radix salvia planted in Zhongjiang, Sichuan province under the good agricultural procedure (GAP).

METHODAfter being prepared, the reference material was digested with 65% HNO3 and H2O2 by microwave-assisted oven systems, and the heavy metals in Radix salvia reference material were accurately determined by inductively coupled plasma mass spectrometry (ICP-MS).

RESULTThe measuring method was validated by running certificated reference materials, including bush twigs and leaves (GBW07602) and tea twigs and leaves (GBW07605) under the same conditions. The recoveries of the elements mostly ranged from 90% to 110%, and the RSD was within 5%. The measurement of radix salvia reference material was carried out by different laboratories with ICP-MS and by several time intervals in one year for the stability. The results showed that the concentration of the heavy metals provided were accurate and the reference material was stable.

CONCLUSIONThe reference material is suitable to be the criterions of heavy metals for radix salvia in the qualities controlling, and is also suitable to be the criterion of poisonous heavy metals of other herbs in the administration of GAP.

Agriculture ; standards ; Arsenic ; analysis ; Cadmium ; analysis ; Drug Stability ; Lead ; analysis ; Metals, Heavy ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; growth & development ; Quality Control ; Salvia miltiorrhiza ; chemistry ; growth & development

Purpose:To study the diagnostic value of using ultrasonography(US)、plain film of the abdomen(KUB)、 intravenous urography (IVU) in acute ureteral calculous obstruction. Methods: The routine B-US、diuretic US、 KUB、 IVU were retrospectively analyzed in 352 cases of acute ureteral calculous obstruction.Results: Of all the patients, 83.2 % (293/352) 、 96.6 % (340/352)were diagnosed as hydronephrosis by routine B-US、 routine B-US plus diuretic US, respectively. The hydronephrotic diagnosis of 59 suspected patients were79.7% (47/59)using diuretic US. The ureteral calculous dignosis of US、KUB、US plus KUB were 76. 1 % (268/352 )、81.6(284/348)and 91.8(323/352),respectively. Conclusion:The routine B-US is an effective method todiagnose acute upper urinary tract obstruction,and routine B-US plus diuretic US is more valuable. B-US plusKUB is the first approach to diagnose ureteral calculi.

OBJECTIVETo investigate the clinical pathological characteristics and prognosis of gastrointestinal stromal tumors (GISTs).

METHODSOne hundred and seven cases, admitted to our hospital from Apr. 1996 to Oct. 2005, were detected by Envision immunohistochemical method and diagnosed as GISTs. Their pathological features, immunohistochemical phenotypes, clinical manifestations and imaging findings were analyzed.

RESULTSOf the 107 GISTs, 107 cases were positive for vimentin (107/107, 100%), 107 cases were positive for CD117 (107/107, 100%), 89 cases were positive for CD34 (89/107, 83.2%), 14 cases were positive for SMA (14/107, 13.1%), 10 cases were positive for desmin (10/107, 9.3%), 22 cases were positive for S-100 (22/87, 20.6%) and 15 cases were positive for NSE (15/107, 14.0%). Among all the GISTs, 73 cases occurred in stomach (68.2%), 28 in small intestine (26.2%), 1 in colon (0.9%) and 5 occurred in other position including mesentery, omentum, and retroperitoneum (4.7%). Fifteen cases were diagnosed as very low grade (14.0%), 25 cases as low grade (23.4%), 33 cases as low malignancy (30.8%) and 34 cases as high malignancy (31.8%). The follow-up was obtained successfully in 89 cases (83.2%). Fourteen cases (13.1%) were confirmed to have recurrences or metastases by review and medical records.

CONCLUSIONSThe diagnosis of GIST depends on pathological observation and immunohistochemical study. CD117 is a sensitive marker for the diagnosis of GIST. Surgical resection is the choice for treating GIST. Extended resection, even combined resection of involved organs, is required for malignant GIST.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastrointestinal Stromal Tumors ; diagnosis ; pathology ; surgery ; Humans ; Immunohistochemistry ; Male ; Middle Aged

OBJECTIVETo investigate the clinical safety and effects of auto-dendritic cells pulsed with HLA-A201-binding peptides prostate-specific antigen (PSA) , prostate-specific membrane antigen (PSMA) and prostatic acid phosphatase (PAP) in the treatment of hormone-refractory metastatic prostate cancer (HRPC).

METHODSSixteen HRPC patients with positive HLA-A201 were enrolled and their monocytes isolated and induced into dendritic cells with the combination of rhGM-CSF and rhIL4. The patients were inoculated subcutaneously near the inguinal region with auto-DCs pulsed with peptides PSA (KLQCVDLHV) , PSMA (ALDVYNGL L) and PAP (LLHETDSAV) every 2 weeks for 4 times, and the immunological and clinical responses were examined within 1 -2 weeks after the final vaccination.

RESULTSVaccination of dendritic cells was well tolerated and no toxicity was observed. The cytokine levels in the serum such as IL-2, IL-12 and IFN-gamma were significantly increased after the vaccination (P < 0.01). The delayed type hyper- sensitivity (DTH) test was positive in 4 of the patients (4/11), the percentage of antigen-special IFN-gamma+ CD8+ T increased in 5 (5/11), the level of the tumor marker PSA decreased in 6 (6/16) , hydrops abdominis reduced in 1 (1/16), and the size of the cervical lymph node lessened in 1 (1/16). Three patients showed partial remission (PR), 7 stability of the disease (SD), and the other 6 progression of the disease (PD).

CONCLUSIONAuto-DC vaccines loaded with PSA, PSMA and PAP peptides, capable of eliciting specific immune responses in HRPC patients, is a safe and effective option for the treatment of advanced HRPC.

Acid Phosphatase ; Aged ; Antigens, Surface ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Cancer Vaccines ; immunology ; Cytokines ; blood ; Dendritic Cells ; immunology ; Glutamate Carboxypeptidase II ; immunology ; HLA-A2 Antigen ; immunology ; Humans ; Male ; Middle Aged ; Prostate-Specific Antigen ; immunology ; Prostatic Neoplasms ; therapy ; Protein Tyrosine Phosphatases ; immunology ; Treatment Outcome