In this paper, metabolic networks of the Corynebacterium glutamicum GWY020 and the two de-rivatives carrying additional mutations HUI821and GUI089 were established and modified. The concentra-tions of extra-cellular metabolites were determined under sub-steady-state (50 h~52 h) of the batch culture. The metabolic flux distribution maps of the three strains were obtained, compared and analyzed. These re-sults indicate that the introduction of analog supersensitive marker or analog resistant marker skew the metabolic flux towards the formation of L-Arginine. This study revealed the usefulness of the metabolic flux analysis as a tool for verification of existing production strains. The analysis may play an important role in helping us to rationally re-design metabolism for further improvement of fermentation process.

Adult ; China ; epidemiology ; Dust ; Humans ; Iron ; Male ; Metallurgy ; Occupational Exposure ; Pneumoconiosis ; diagnostic imaging ; epidemiology ; prevention & control ; Radiography ; Risk Factors ; Steel ; Welding

Objective To evaluate cerebrospinal fluid adenosine deaminase(CSF-ADA)activity in the diagnosis of tuberculous meningitis(TMB), and to observe its dynamic changes. Methods A total of 160 patients were included and were divided into two groups: 76 cases of TBM and 84 cases of non-TBM.Among the cases of non-TBM, there were 36 cases of bacterial meningitis, 30 cases of viral meningitis and 18 cases of cryptocoocal meningitis. All the patients were measured with their CSF-ADA activity by Enzymecoupled assay(Trinder method)and 47 patients of TBM were measured again after 2 weeks' and 6 weeks'antitubercular therapy. Results were expressed as(-x)± s. Mann-Whitney U test and paired-samples t test were used. Results CSF-ADA activity in TBM group was(12.9 ±6.4)U/L, while that in the non-TBM group was(6.0 ± 4.1)U/L, the difference was of statistical significance(U = 7.860, P ＜ 0.05). With the cutoff value of 9 U/L, the sensitivity and specificity to differentiate TBM from non-TBM was 84.21% and 83. 33%, respectively. CSF-ADA activity decreased in TBM patients after antitubercular treatment.Conclusions CSF-ADA activity can be an effective laboratory marker for early differential diagnosis of TMB with the cut-off value of 9 U/L. Dynamic changes of CSF-ADA activity may be a indicator for the effect of antitubercular treatment.

Animals ; Cells, Cultured ; Hepatocytes ; drug effects ; secretion ; Male ; Pancreatic Polypeptide ; pharmacology ; Rats ; Rats, Sprague-Dawley

The diagnosis and treatment of prostate cancer are being improved due to the popularized screening of prostate specific antigen. Advanced prostate cancer, in spite of its response to androgen deprivation therapy, may finally develop into castration-resistant prostate cancer (CRPC) and shorten the overall survival of the patients. Many efforts have been made by worldwide researchers for new approaches to the management of CRPC, including new hormonal therapy, cytotoxic chemotherapy, immunotherapy, and bone metastasis-targeted therapy. This paper reviews the emerging agents undergoing clinical evaluation and drugs that have received approval for the treatment of CRPC in order to provide doctors and patients with more treatment options for CRPC and improve the overall survival rate and quality of life of the patients.
Androgen Antagonists ; Bone Neoplasms ; prevention & control ; Humans ; Immunotherapy ; Male ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms, Castration-Resistant ; therapy ; Quality of Life

Objective To investigate the clinical characteristics of hypertrophic cardiomyopathy (HCM) in the elderly. Methods The clinical characters of HCM in 70 elderly patients were retrospectively analysed. Results Among the 196 identified HCM patients, 70 were elders. Out of them, 7 patients(10.0%) were suspected as HCM according to the clinical symptoms, 29 patients (41.4%)were suspected as other cardiac diseases, 34 (48.3%) were diagnosed HCM due to other reasons. Among the 70 patients, 12 patients(17.1%) had history of cerebrovascular diseases, 54 (77.1%) manifested symptoms after 45 years of age and 4 (5.7%) showed no obvious symptoms. Among them, 18 patients were examined by UCG for two times and HCM was diagnosed at the second time. Conclusions HCM in the elderly is not an uncommon disease. The onset of cardiac symptoms is relatively late in the elderly HCM and frequently misdiagnosed; cerebrovascular diseases are commonly seen in elderly HCM patients.

To explore the protective effect of sodium tanshinone IIA sulfonate (STS) on microcirculatory disturbance of small intestine in rats with sepsis, and the possible mechanism, a rat model of sepsis was induced by cecal ligation and puncture (CLP). Rats were randomly divided into 3 groups: sham operated group (S), sepsis group (CLP) and STS treatment group (STS). STS (1 mg/kg) was slowly injected through the right external jugular vein after CLP. The histopathologic changes in the intestinal tissue and changes of mesenteric microcirculation were observed. The levels of tumor necrosis factor-α (TNF-α) in the intestinal tissue were determined by using enzyme-linked immunoabsorbent assay (ELISA). The expression of intercellular adhesion molecule-1 (ICAM-1) in the intestinal tissue was detected by using immunohistochemisty and Western blot, that of nuclear factor κB (NF-κB) and tissue factor (TF) by using Western blot, and the levels of NF-κB mRNA expression by using RT-PCR respectively. The microcirculatory disturbance of the intestine was aggravated after CLP. The injury of the intestinal tissues was obviously aggravated in CLP group as compared with S group. The expression levels of NF-κB p65, ICAM-1, TF and TNF-α were upregulaed after CLP (P<0.01). STS post-treatment could ameliorate the microcirculatory disturbance, attenuate the injury of the intestinal tissues induced by CLP, and decrease the levels of NF-κB, ICAM-1, TF and TNF-α (P<0.01). It is suggested that STS can ameliorate the microcirculatory disturbance of the small intestine in rats with sepsis, and the mechanism may be associated with the inhibition of inflammatory responses and amelioration of coagulation abnormality.

Objective To study the clinicopathologic characteristics and differential diagnosis of T-cell immunophenotype in intestinal non-Hodgkin's lymphoma(NHL).Methods The clinicopathologic characteristics of 13 cases with intestinal T-cell lymphoma were analyzed by light microscopy and immunohistochemistry(Envision detection method).Results The lesions of 8 cases with T-cell lymphoma were found on the small intestine and 5 on the colon.Grossly,8 cases showed ulcer pattern,3 polypoid pattern and 2 presented as a regional thickening of intestinal wall.The tumor cells were medium to large size with pleomorphic nuclei and inflammatory background.The neoplastic lesions expressed the immunophenotype of peripheral T cells.The neoplastic cells of 13 cases(100%)expressed leukocyte common antigen(LCA);10(76.9%)cases expressed CD3;9(69.2%)CD45RO;5(38.5%)EB virus(EBV);3(23.1%)CD56 and 2(15.4%)vimentin(VIM).All the cases were negative for CD20,CD79a,CK,CDX2,NSE,CgA and CD117.ConclusionIntestinal T-cell lymphoma is a rare,aggressive neoplasm with poor prognosis and should be distinguished from other malignant tumors of intestine.

OBJECTIVETo construct and identify a lentiviral vector of RNA interference targeting human Notch-1 gene.

METHODSTo determine the Notch-1 gene sequences, three RNAi target sequences (shRNA1-3) were designed in accordance with the RNAi sequence design principles and cloned into the lentiviral vector pLenOR-THM by endonuclease BamH I restriction, EcoR I double digestion, and T4 DNA-ligase ligation. After the transformation into competent DH5alpha bacteria, the candidate clones were identified by Kpn I and EcoR I double digestion and DNA sequencing. The recombinant and three packaging plasmids were co-transfected into human embryonic kidney cell line 293T cells by lipofectamine to produce the lentiviral particles. The viral titer was determined. The 293T cells were infected by the lentiviral particles obtained, and transfection efficiency was assessed using a fluorescent microscope. The lentiviral vector particles were also transfected into ACC-M cells. The Notch-1 expression in the transfected cells was assayed by quantitative reverse transcription polymerase chain reaction (QRT-PCR) and Western blot analysis.

RESULTSThe lentiviral RNAi vector pLenOR-THM-Notchl for Notch-1 gene was constructed successfully. Strong green fluorescence was observed in the 293T cells under fluorescent microscope after co-transfection of the cells with the four plasmids of lentiviral vector. The virus in the supernatant reached a titer of 5.8 x 10(8) TU x mL(-1). The transfection efficiency of the collected virus exceeded 90% in 293T cells with 1 as a multiplicity of infection. The third lentiviral vector was found to significantly inhibit the Notch-1 expression at the mRNA and protein levels.

CONCLUSIONThe lentiviral RNAi vector of Notch-1 has been successfully constructed and identified.

Cell Line ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Receptor, Notch1 ; Transfection

Objective To observe the effect of Ti-6AL-4V particles on morphological and func- tional changes of osteoclast in vitro.Methods Mature osteoclasts separated from New Zealand Rabbits were cultured on glass slices and cortical bone slices.The experimental group was stimulated by Ti-6AL- 4V particles at concentration of 0.1 mg/ml.The cells were stained with TRAP at different culture time to observe the morphological variety.The bone resorption pits on bone slices were stained by toluidine blue and the resorption areas analyzed by computer image analysis software.Results Osteoclasts phagocy- tosed the particles,with irregular shapes,deeper TRAP stain and earlier apoptosis.Stimulation by Ti- 6AL-4V particles brought about larger area of bone absorption lacuna.Conclusion Osteoclasts have the ability to phagocytose Ti-6AL-4V particles,which leads to morphological and functional changes and enhances bone resorption.