Mycena subpiligera, a new taxon in sect. Fragilipedes that can strongly enhance the germination efficiency of Gastrodia elata seeds, was discovered in subtropical areas of China. As revealed by a morphological comparison with related Mycena species as well as maximum likelihood (ML) and Bayesian phylogenetic analyses based on sequences of the internal transcribed spacer (ITS) and the large subunit (LSU) regions of nuclear ribosomal RNA, the new taxon can be distinguished from phenotypically similar and phylogenetically related species. Optimal cultural conditions for M. subpiligera basidiomata are reported, and the germination rate of the new species is compared with that of M. citrinomarginata.

BACKGROUND AND OBJECTIVEThe basic structure of salicylaldehyde-amino acid Schiff base compounds includes a C=N chemical bond. These compounds show significant antitumor activities in vitro when combined with a metal ion. This study investigated the effects and possible mechanisms of four salicylaldehyde-amino acid Schiff base copper ternary coordination compounds on the proliferation of human gastric cancer cell line BGC823.

METHODSThe BGC823 cells were treated with the four compounds (6B, 7B, 6P, and 7P). Cell proliferation was detected by MTT assay. Apoptosis and changes in the cell cycle were analyzed by flow cytometry. DNA damage was observed using a DNA ladder assay. The expression of p53 protein was determined by immunocytochemistry.

RESULTSThe proliferation of BGC823 cells was significantly inhibited by the four compounds and the effect was concentration-dependent. The half maximal inhibitory concentration (IC50) of 6B, 7B, 6P, and 7P for BGC823 cells were 18.10, 27.50, 3.61, and 3.45 micromol/L, respectively. Flow cytometry showed the four drugs induced apoptosis in BGC823 cells, which was confirmed by DNA ladder experiments. Flow cytometry also detected changed phases in the cell cycle from treatment with the compounds. The percent of cells in the G(0)/G(1) phase decreased and that of cells in the G1/S and G(2)/M phases increased, indicating that S-and G2-phase blockages exist. As shown by immunocytochemistry, the expression of p53 decreased in BGC823 cells treated with the four drugs, indicating the involvement of the p53 pathway to BGC823 cell apoptosis.

CONCLUSIONSThe four compounds showed significant activities on restraining proliferation of BGC823 cells in vitro, induced apoptosis, and caused changes in the cell cycle. This may be related to the downregulation of p53.

Aldehydes ; chemistry ; Amino Acids ; chemistry ; Antineoplastic Agents ; chemical synthesis ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Coordination Complexes ; chemical synthesis ; pharmacology ; Copper ; chemistry ; Humans ; Inhibitory Concentration 50 ; Schiff Bases ; chemistry ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism

Tropane alkaloids (TAs) are a class of anticholinergic drugs widely used in clinical practice and mainly extracted from plant, among which Atopa belladonna is the main commercial drug source. It is of great industrial value to obtain TAs in large quantities by plant metabolic engineering. In TAs pathway, cytochrome oxidase CYP82M3 catalyze the synthesis of tropinone and then tropinone reductase I (TRI) compete with TRII for tropinone to form tropine leading to the TAs synthesis (drainage). In this study, based on the "increasing flow and drainage" metabolic engineering strategy, two genes, namely HnCYP82M3 and DsTRI from Hyoscyamus niger and Datura stramonium, respectively, were overexpressed in the hair roots of A. belladonna, with a view to promote the TAs accumulation. The HnCYP82M3 gene was cloned from the root of H. niger, and it encoded amino acid with 91.7% sequence identity with AbCYP82M3 from A. belladonna. Overexpression of HnCYP82M3 alone did not affect the content of TAs in hair roots of A. belladonna, indicating that CYP82M3 was not a key enzyme in TAs biosynthesis. Simultaneous overexpression of HnCYP82M3 and DsTRI greatly promoted the accumulation of the three TAs, and the contents of hyoscyamine, anisodamine and scopolamine were 4.97 times, 2.83 times and 2.19 times that of the control, respectively, and the increase amplitude was greater than that of single overexpression of DsTRI. This study showed that the "increasing flow and drainage" strategy of enzyme genes co-expression at branch points was a promising metabolic engineering method to effectively improve the biosynthesis of TAs in A. belladonna, and laid a theoretical and technical foundation for the large-scale industrial acquisition of TAs.

The rol genes on pRiA4 plasmid of Agrobacterium rhizogenes are potent genes that promote secondary metabolism. Molecular breeding of Atropa belladonna can be conducted by introducing rol genes to increase tropane alkaloids (TAs) content in A. belladonna. In this study, the rolB gene was overexpressed in A. belladonna plants to study the effect of rolB gene on the biosynthesis of TAs. The phenotype, TAs content and expression levels of key enzyme genes in the pathway of TAs biosynthesis of transgenic A. belladonna were analyzed. The results showed that transgenic A. belladonna had developed root system, enlarged leaves, increased leaf fresh weight, deepened leaf color, enlarged flowers, changed flower shape, reduced pistil height and decreased pollen vitality. The content of TAs in the stems of transgenic A. belladonna was significantly higher than that of the control, and the contents of scopolamine, anisodamine, hyoscyamine can reach 2.11-2.91, 1.23-2.37 and 4.88-5.20 times of the control, respectively. Compared with the control group, the expressions of key enzymes putrescine N-methyltransferase (PMT), type III polyketide synthase (PYKS), tropinone reductase I (TRI), aromatic amino acid aminotransferase 4 (ArAT4), UDP-glycosyltransferase 1 (UGT1) and hyoscyamine 6-β-hydroxylase (H6H) in the TAs biosynthesis pathway were up-regulated, and the expression of tropinone reductase II (TRII) as a metabolic shunting gene was down-regulated. The results indicated that rolB gene enhanced TAs synthesis ability in roots and accumulation in stems of A. belladonna by enhancing metabolic flow of TAs synthesis pathway and weakening the metabolic shunt of competing pathway. This study laid a foundation for molecular breeding of A. belladonna with high-yield TAs content using rolB gene.

OBJECTIVETo study the factors affecting the management and treatment of immigrant workers with tuberculosis, to determine the most effective measures and to provide an academic basis for tuberculosis control and prevention strategies targeting the immigrant population.

METHODSA self-designed questionnaire was administered to 1364 immigrant workers with tuberculosis (study group) and 436 local residents (control group) with tuberculosis.

RESULTSWhether a patient's tuberculosis status was discovered or not was related to the medical facilities initially visited, job mobility, and the individual worker's economic condition. The percentage of cases discovered was relatively low among those who made their first visit to a private clinic (39.0%), and 58.4% (796) of immigrant workers delayed their first consultation mainly due to neglecting symptoms (55.9%, 445), subjective perception of symptoms being not severe enough (19.3%, 154) to visit a physician and the inconvenience of visiting a hospital because of its being far away from home (15.8%, 47). As comparing immigrant workers with local residents, the differences of the delay reason between the two groups were statistically significant (chi(2) = 21.49, P < 0.01). And 39.9% (544) of immigrant workers and 49.1% (214) of local residents had had late confirmation. The differences of the delay reasons between the two groups were insignificant (chi(2) = 7.31, P = 0.293). And 17.6% (240) patients of immigrant workers and 13.1% (57) patients of local residents did not keep to their drug regimens in a timely fashion. The differences of the reasons between the two groups were insignificant (chi(2) = 6.66, P > 0.05). And 77.5% (1057) of immigrant workers and 31.8% (138) of local residents were considered that taking medicine in supervision spot might have impact on their lives and work. The differences of the reasons between the two groups were significant (chi(2) = 9.71, P < 0.05). All 79.2% (1080) of immigrant workers and 63.3% (276) of local residents did not obtain medicine according to prescriptions. The differences of reasons between the two groups were statistically significant (chi(2) = 24.84, P < 0.01). And 51.2% (699) of immigrant workers and 46.1% (201) of the registered population did not follow up with doctors' directions for lab tests. The differences of the reasons between the two groups were insignificant (chi(2) = 3.26, P > 0.05).

CONCLUSIONThe influential factors in tuberculosis management and treatment were complex. Prevention strategies should focus on health education and promotion activities to improve awareness in seeking medical services. Also, developing and standardizing reference mechanisms for patients, and alleviating the economic burden of the workers will be critical to reduce the tuberculosis incidence.

China ; epidemiology ; Factor Analysis, Statistical ; Humans ; Transients and Migrants ; Treatment Outcome ; Tuberculosis, Pulmonary ; epidemiology ; prevention & control ; therapy ; Urban Population

OBJECTIVETo study the expression of lung Krüppel-like transcription factor (KLF2/LKLF) in lung tissues of rats with chronic obstructive pulmonary disease (COPD) and the relationship between KLF2 and NF-E2-related factor 2 (Nrf2), and make further explore the effects of KLF2 on the expression of gamma-glutamylcysteine synthetase (gamma-GCS).

METHODSTwenty-two male SD rats were randomly divided into a COPD group (n = 10) and a normal control group (n = 11). The rat model of COPD established by cigarette smoking and intratracheal instillation of lipopolysaccharide (LPS), and lung tissues were obtained. The expressions of KLF2, Nrf2, gamma-GCS mRNA and protein in lung tissues were measured by immunohistochemistry (IHC), Western blot, in situ hybridization (ISH) and reverse transcription-polymerase chain reaction (RT-PCR). To explore the relationship between KLF2 and Nrf2 protein,we utilize the method of co-immunoprecipitation (CO-IP).

RESULTSIHC and Western blot showed that protein expressions of KLF2, Nrf2, gamma-GCS were higher in the lung tissues from rats with COPD than those in the control groups (all P < 0.05). The levels of KLF2, gamma-GCS mRNA were markedly increased in the COPD group (all P < 0.01) while Nrf2 mRNA expression in COPD group had no significant difference with that in control group ( P > 0.05). CO-IP result showed that KLF2 were obviously present in immunoprecipitates of Nrf2 (P < 0.01) . Linear correlation analysis showed that the level of KLF2 protein was positively correlated with the level of Nrf2 protein (P < 0.05), and KLF2, Nrf2 proteins were positively correlated with gamma-GCS mRNA and protein (all P < 0.05).

CONCLUSIONThe expression of KLF2 is significantly up-regulated in COPD, which maybe up-regulate gamma-GCS mRNA expression by increasing Nrf2 expression and nuclear translocation against oxidative stress.

Animals ; Dipeptides ; metabolism ; Kruppel-Like Transcription Factors ; metabolism ; Lung ; pathology ; physiopathology ; Male ; NF-E2-Related Factor 2 ; metabolism ; Pulmonary Disease, Chronic Obstructive ; metabolism ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the safety and feasibility of the retrosigmoid suprameatal approach (RSSMA) for petrous apex resection. Methods Ten human dry skull and 18 cadaverie skull specimens were collected and 3-dimensional CT scanning was performed with slice thickness of 1 mm. Craniotomy was performed on the specimens through a modified retrosigmoid approach, and the suprameatal tubercle (ST) and petrous apex (PA) were removed without damaging the trigerninal and facial nerves. The petrous bone was resected to the farthest lateral margin (FLM) that the approach could allow. CT-based and manual measurements were used to determine the lateral-middle line, superior-inferior, anterior-posterior lengths of the ST and PA. The superolateral lip of the internal auditory meatus (SLIAM) was defined as the landmarks for the measurement, and the distances from the SLIAM to the fundus, the common crus, and vestibule was determined. Results From thesuperior-inferior to the anterior-posterior and median-lateral directions, the resection rate of the PA increased to (26.6±6)%, (45±5)%, and (72±6)%, and the rate for the ST to (69±10)%, 100%, and 100%, respectively. The resection rate of the PA at the siphonal portion was (44±7)%. In the RSSMA, the distance from the SLIAM to the FLM (17.6±2.0 mm) was greater than the distances from the SLIAM to the vestibule (10.1±1.4 mm), the fundus (10.4±1.5 mm), and the common crus (10.6±1.1 mm). Conclusions The RSSMA may well protect the siphonal portion of the internal carotid artery from damages in PA resection. The FLM of the RSSMA is always lateral to the vestibule and the fundus of the internal auditory canal and the common crus, and therefore injuries to the vestibule, the semicircular canal and the common crus should be avoided.

OBJECTIVETo compare therapeutic effects of Tongdu Tiaojin needling method and routine acupuncture method on blepharoptosis.

METHODSFifty-eight cases of blepharoptosis were randomly divided into an observation group and a control group, 29 cases in each group. The observation group was treated by Tongdu Tiaojin needling method with Baihui (GV 20), Fengchi (GB 20), Houxi (SI 3), Yintang (EX-HN 3) selected, and the control group by routine acupuncture method with Cuanzhu (BL 2), Yangbai (GB 14), Sizhukong (TE 23) selected. They were treated for 3 courses and one course was constituted by 10 sessions.

RESULTSThe cured rate of 41.4% and the effective rate of 82.8% in the observation group were better than 24.1% and 48.2% in the control group (P<0.05); after treatment, the palpebral fissure widened and the level of the palpebra superior shading the cornea was alleviated in the two groups, but the observation group was superior to the control group (P<0.05).

CONCLUSIONTongdu Tiaojin needling method has a definite therapeutic effect on blepharoptosis.

Acupuncture Points ; Acupuncture Therapy ; methods ; Adult ; Blepharoptosis ; physiopathology ; therapy ; Eye ; physiopathology ; Female ; Humans ; Male

OBJECTIVETo isolate the wild-type virulent phage of Helicobacter pylori (Hp) and simulate the treatments in vitro to investigate the methods for oral Hp-assisted penetration of the phage through the gastric barrier and offspring phage release for infection and treatment of gastrointestinal Hp.

METHODSThe Hp strain was cultured with the candle cylinder method and the virulent phage was isolated by single plate or double plate experiment. A simulated gastric juice was applied and the bactericidal effect of the phage was tested with double flats experiment.

RESULTSAfter a 1.5-h treatment in simulated gastric juice, the orally derived Hp-borne phage was still capable of forming plaques while the control phage was not.

CONCLUSIONThe oral Hp can help the phage resist the gastric juice and then infect the gastrointestinal Hp.

Bacteriophages ; isolation & purification ; physiology ; Gastrointestinal Tract ; microbiology ; Helicobacter Infections ; therapy ; Helicobacter pylori ; virology ; Humans ; Virulence

OBJECTIVETo evaluate the disadvantage of nonstandard operation for thyroid papillary cancer, and the value of re-operation.

METHODSEight hundred and ninety three thyroid papillary cancer patients (332 patients who received nonstandard operation received re-operation and 561 patients who received first standard operation in Zhejiang Cancer Hospital from January 1990 to January 2000) were retrospectively reviewed.

RESULTSPathological results confirmed that there were 53.9% cases with residual cancer in re-operative specimen. The parathyroid was preserved in 74.1% cases in re-operation and 93.0% cases in standard operation (P < 0.01). The recurrent laryngeal nerve was injured in 3.3% cases in reoperation and 1.2% cases in standard operation (P < 0.05). The positive neck lymph node was 39.2% and 37.4% (P > 0.05), the 5-year local recurrence rate was 7.5% and 3.7% (P < 0.01), the total 5-year and 10-year cum-survival rates were 90.2%, 84.4% in patients with re-operation and 94.0%, 92.5% in patients with standard operation. The 10-year cum-survival rate were 67. 8% in patients with recurrence and 92. 9% in patients with non-recurrence in primary site respectively. Statistical analysis showed that the cancer recurrence influences the survival rate significantly (P < 0.01).

CONCLUSIONSIn consideration of the higher residual tumor rate in patients who received nonstandard operations, the re-operations were necessary. But the re-operation could also leads to higher recurrence rate and more functional injuries. The standardized primary operation should therefore be formulated and advocated.

Adolescent ; Adult ; Aged ; Carcinoma, Papillary ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Reoperation ; Thyroid Neoplasms ; surgery ; Thyroidectomy ; methods ; Young Adult