According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, a full-length cDNA sequence of SQS2 from S. miltiorrhiza f. alba was cloned by the method of reverse transcription polymerase chain reaction (RT-PCR). The SmSQS2 cDNA sequence was obtained, this sequence is named SmSQS2 and its GenBank registration number is KM244731. The full length of SmSQS2 cDNA was 1245 bp, encoding 414 amino acids including 5'UTR 115 bp and 3'UTR 237 bp. Sequence alignment and phylogenetic analysis demonstrated that SmSQS2 had relative close relationship to the SQS2 of S. miltiorrhiza. The induction of E. coli [pET28-SQS2] in different temperature, induction time, IPTG concentrations and density of inducing host bacterium (A600) were performed, Shaking the culture at 30 degrees C until the A600 is approximately 0.6 and add IPTG to final concentration of 0.2 mmol x L(-1), and then the optimal expression of SmSQS2 recombinant protein were accumulated after the induction time of 20 h. The research provided important base for the study of sterol and terpene biosynthesis of SQS2 in S. miltiorrhiza f. alba.
Cloning, Molecular ; Farnesyl-Diphosphate Farnesyltransferase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; classification ; enzymology ; genetics ; Sequence Alignment

Background and purpose: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a new minimally invasive method in the dignosis for mediastinal lymph nodes. This study was to evaluate the diagnostic yield of EBUS-TBNA for mediastinal lymph nodes. Methods: Twenty patients with mediastinal lymph nodes found by CT underwent the dignosis by EBUS-TBNA form April 1st 2009 to July 16th 2009. The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of EBUS-TBNA were evaluated. Results: Twenty patients with 37 lymph node groups were studied. Overall sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of EBUS-TBNA for diagnostic were 84.62%, 100%, 100%, 77.78% and 90.00%, respectively. The diagnostic accuracy for cancer was 100%. The operation time was 11.9min per group in average with no serious complication. The median length of hospital stay was 1 (range from 1 to 17 days) day after operation. There were significant differences in the average operation time between the first three patients and the others (36.25 min vs. 7.76 min; z=3.247, P=0.001). Conclusion: Endobronchial ultrasound-guided transbronchial needle aspiration proved to be a safe procedure with a high yield for the diagnosis of mediastinal lymph nodes.

OBJECTIVETo assess the potential impact of warming climate in winter to the scale and severity of schistosomiasis epidemics in China.

METHODSBased on the data of mean temperature and monthly minimum temperature in January from 126 (out of 733) weather observation stations in China, the trend and oscillation amplitude was analyzed. The impact of warming climate in winter to the scale of schistosomiasis spreading was assessed, using the indices of 0 degrees C mean temperature and -4 degrees C mean monthly minimum temperature in January. Correlation between these two indices was analyzed.

RESULTSEvidences showed the warming climate in winter was seen all over China with the mean monthly minimum temperature and the mean temperature in January had increased 1.3 degrees C and 0.9 degrees C since 1986. The contours of mean monthly minimum temperature -4 degrees C and mean temperature 0 degrees C in January moved 1 - 2 latitudes northward.

CONCLUSIONThe warming climate phenomenon seen in winter was considered favorable to the living of snails in winter, resulting in the possible increase of northward spreading of snails.Thus, monitoring of snails should be paid special attention.

Animals ; China ; epidemiology ; Climate ; Disease Reservoirs ; Female ; Health Education ; Humans ; Male ; Prevalence ; Schistosoma japonicum ; physiology ; Schistosomiasis japonica ; epidemiology ; prevention & control ; Seasons ; Snails ; parasitology ; physiology ; Temperature

OBJECTIVE: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the determination of opiates in biological samples according to the emerging problem in drugs abuse. METHODS: Opiates such as heroin, 6-acetylmorphine, morphine, codeine, acetylcodeine, hydrocodone and hydromorphone were isolated from human blood, urine, oral fluid and hair using a simple extraction and consequently analyzed using LC-MS/MS. The method was evaluated by real cases. RESULTS: The mobile phase give the optimum separation for opiates. The detection limit of morphine in urine with dilution and liquid-liquid extraction and in hair is 10ng/mL, 0.01 ng/mL and 0.01 ng/mg, respectively. CONCLUSION The method is simple and rapid, offering superior sensitivity and selectivity for opiates. The target compounds comprising hydrocodone and hydromorphone enlarge the applied area.
Chromatography, Liquid ; Codeine/analysis* ; Forensic Medicine/methods* ; Hair/chemistry* ; Humans ; Hydrocodone/analysis* ; Hydromorphone/analysis* ; Morphine/analysis* ; Narcotics/analysis* ; Reproducibility of Results ; Saliva/chemistry* ; Substance Abuse Detection/methods* ; Tandem Mass Spectrometry

Objective To investigate diffusion-weighted(DWI)magnetic resonance imaging (MRI) features of the thrombi in patients with cerebral sinus thrombosis(CST),and explore their relation with the recanalization rate alter thrombolytic therapy. Methods The clinical and imaging data were retrospectively analyzed in 14 CST cases,including conventional MRI,FLAIR,DWI and MRV features.The hyperintense signals on DWI at the site ofvenous occlusion were analyzed,and MRV was performed 1-3 months after the thrombolytic therapy to assess the recanalization.The relation between the findings by T1WI,FLAIR and DWI before the thrombolytic therapy and the recanalization rate after the therapy was evaluated.Results Hyperintense signals Was detected in 21 occluded sinuses in 9 patients(41%)with a recellt CVT.Hyperintense signals in the thrombi on the initial MRl were associated with a significantly lowered rate of complete recanalization of the vessel as compared with that among cases without the hyperintense signals(19.0%vs 68.2%,P=0.005). Conclusion The DWI signals of the thrombi in the cerebral venous sinuses can be associated with the progression of the thrombosis,and the presence of hyperintense signals in the occluded cerebral sinuses on the initial DWI may suggest a low rate of sinus recanalization after 1 t0 3 months of thrombolytic therapy.

OBJECTIVETo investigate the effects of host-derived p38 mitogen-activated protein kinase subunit 38 (p38MAPK) and the hepatitis B virus X antigen (HbxAg) on cell proliferation and apoptosis in human hepatocellular carcinoma (HCC), and to study the mechanism underlying hepatocarcinogenesis.

METHODSLiver tissues were biopsied from healthy individuals and patients with chronic hepatitis B (CHB), liver cirrhosis, paratumor cirrhosis, and HCC. Immunohistochemical staining was used to detect expressions of HBxAg, p38MAPK, cell cycle G2/M phase-related factors (cdc25B, p34cdc2, cyclin B1), and cell proliferation factor ki-67.The terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling method (known as TUNEL) was used to detect apoptosis.

RESULTSThe highest rates of HBxAg were detected in CHB (65.0%) and HCC (44.4%) liver samples, and the antigen was mainly expressed in nuclei. Increasingly higher rates of p38MAPK, cdc25B, cyclin B1, and p34cdc2 expression were detected with increases in disease severity: normal liver (40.0%, 20.0%, 20.0%, and 30.0%, respectively), chronic hepatitis B (60.0%, 65.0%, 40.0%, and 50.0%, respectively), liver cirrhosis (65.0%, 75.0%, 70.0%, and 55.0%, respectively), paratumor cirrhosis (66.7%, 75.0%, 75.0%, and 63.9%, respectively), and HCC (77.8%, 80.6%, 80.6%, and 72.2%, respectively). In addition, the intracellular location of p38MAPK expression was different under different disease conditions, showing nuclear expression in CHB and liver cirrhosis samples and cytoplasmic expression in paratumor cirrhosis and HCC samples (x2 = 1.11, P more than 0.05). The proliferation index (PI) and the apoptosis index (AI) were both increased along with disease severity (normal more than CHB more than paratumor cirrhosis more than HCC) (PI: 0.0000+/-0.000, 0.0502+/-0.011, 0.0411+/-0.009, 0.0762+/-0.017; AI: 0.0351+/-0.024, 0.0607+/-0.022, 0.0562+/-0.013, 0.0716+/-0.011), with the notable exception for liver cirrhosis (PI: 0.1810+/-0.036 and AI: 0.1200+/-0.018). PI in poorly-differentiated HCC (0.2285+/-0.062) was significantly higher than in well-differentiated HCC (0.1216+/-0.032, t = 2.082, P = 0.044). AI in well-differentiated HCC (0.152+/-0.026) was significantly higher than in poorly-differentiated HCC (0.081+/-0.022, t = 2.129, P = 0.041).

CONCLUSIONSIn the process of hepatocarcinogenesis, HBxAg may cause a series of abnormal changes in cell cycle, proliferation and apoptosis by affecting the expression of p38MAPK.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Cycle ; Cell Division ; Cell Proliferation ; Hepatitis B, Chronic ; pathology ; Humans ; Liver Cirrhosis ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Trans-Activators ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

The objective of this study was to evaluate the etiological factors, diagnosis and therapy of pulmonary fungal infection in hematological malignancies, 14 cases of malignant hematological disease with pulmonary fungal infection were collected and analyzed. The results showed that 11 out of 14 cases had the respiratory manifestations, sputum was sticky and difficult to be expectorated in 7 cases, X rays in chests showed shadows with features of stigma and sheet in 11 cases, Candida albicans and aspergillus infection were observed in 10 and 2 cases respectively, the numbers of neutrophil were below 0.5 x 10(9)/L in 8 cases and below 1.0 x 10(9)/L in 3 cases respectively, fluconazole was used for 12 cases and clinical symptoms of 11 cases were improved within two weeks. In conclusion, the occurrence of pulmonary fungal infection in malignant hematological diseases is associated with intensive chemotherapy, decrease of neutrophil counts and using of broad-spectrum antibiotics, the diagnosis at early stage is difficult and clinicians should pay more attention to its clinical and laboratory examinations, and give them therapy in time.
Adolescent ; Adult ; Aged ; Antifungal Agents ; therapeutic use ; Aspergillosis ; complications ; diagnosis ; drug therapy ; Candidiasis ; complications ; diagnosis ; drug therapy ; Female ; Fluconazole ; therapeutic use ; Hematologic Neoplasms ; drug therapy ; etiology ; pathology ; Humans ; Lung Diseases, Fungal ; complications ; diagnosis ; drug therapy ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVEMultilocular brain abscess in children is a serious neurosurgical emergency and remains a serious, life-threatening disease. This study evaluated the role of neuroendoscopy in treating multilocular brain abscess in children.

METHODSBetween January 2002 and June 2007, 16 children with multilocular brain abscess underwent an operation using a pure endoscopic procedure.

RESULTSIncreased intracranial pressure was relieved after operation in the 16 patients. CT/MRI after operation showed the abscess cavities disappeared and only the residual abscess walls existed in the 16 patients. Fourteen patients were followed up for 6 months to 5 years after surgery. Abscess walls disappeared in 13 patients and abscess recurred only in 1 patient.

CONCLUSIONSNeuroendoscopy for treatment of multilocular brain abscess is safe and effective in children.

Adolescent ; Anti-Bacterial Agents ; therapeutic use ; Brain Abscess ; surgery ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Neuroendoscopy ; methods

BACKGROUNDLumiracoxib is a highly selective cyclooxygenase-2 (COX-2) inhibitor with antiinflammatory, analgesic and antipyretic activities comparable with class specific drugs, but with much improved gastrointestinal safety. No studies have examined lumiracoxib for antitumorigenic activity on human nonsmall cell lung cancer cell lines in vitro or its possible molecular mechanisms.

METHODSThe antiproliferative effect of lumiracoxib alone or combined with docetaxol on A549 and NCI-H460 lines was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Drug-drug interactions were analyzed using the coefficient of drug interaction (CDI) to characterize the interactions as synergism, additivity or antagonism. Morphological changes were observed by acridine orange fluorescent staining. Extent of apoptosis was determined by flow cytometry.

RESULTSLumiracoxib (15 - 240 micromol/L) has an inhibitory effect on the proliferation of A549 and NCI-H460 cell lines in concentration- and time-dependent manners with the IC50 values of 2597 micromol/L and 833 micromol/L, respectively. The synergistic effect was prominent when lumiracoxib (15 - 240 micromol/L) was combined with docetaxol (0.2 - 2 micromol/L) (CDI < 1). Fluorescent staining showed that lumiracoxib could induce apoptosis in A549 and NCI-H460 cells. Lumiracoxib treatment also caused an increase of the sub-G1 fraction in each cell line and resulted in an increase of G0/G1-phase cells and a decrease of S-phase cells.

CONCLUSIONSLumiracoxib had antiproliferative effect on the human nonsmall cell lung cancer cell lines A549 and NCI-H460 and had a significant synergy with docetaxol, which may be related to apoptotic induction and cell cycle arrest.

Apoptosis ; drug effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Diclofenac ; analogs & derivatives ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Taxoids ; pharmacology

OBJECTIVETo investigate the prevalence and subtypes of microdeletions in azoospermia factor (AZF) region in infertile men from Sichuan in order to correlate genotypes with phenotypes.

METHODSMultiplex-PCR was used to detect sequence tagged sites (STS) of AZF microdeletions in 1011 infertile men including 713 cases of non-obstructive azoospermia and 298 cases of severe oligospermia.

RESULTSThe overall prevalence of microdeletions was 10.48% (106/1011), and the deletion rates were 11.08% (79/713) in non-obstructive azoospermia and 9.06% (27/298) in severe oligospermia. Complete AZFa or AZFb deletions were associated with azoospermia, whereas AZFc deletion (60.38%) was the most frequent deletion. The deletions were associated with variable spermatogenic phenotypes, and 37.50% of the patients with a deletion had sperms in the ejaculate. A mild decline in sperm concentration was found in two cases with partial AZFb deletion and one case with partial AZFb-c deletion.

CONCLUSIONDeletions of the AZFc region were most commonly found in our patients. All cases with complete AZFa or AZFb deletions and a proportion of cases with AZFc deletion were associated with azoospermia. Our study has provided more insight into the genotype-phenotype correlation, and confirmed that Yq microdeletion screening has a significant value for the diagnosis for male infertility.

Adult ; Azoospermia ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; Genetic Association Studies ; methods ; Humans ; Infertility, Male ; genetics ; Male ; Phenotype ; Young Adult