The genetic diversity of three Tibetan herbs, i. e., Sang-Di, E-Dewa and Ye-Xingba (Tibetan names), was studied based on the field collection, specimen identification and DNA sequence analysis. Swertia hispidicalyx, Gentiana lhassica and Scrophularia dentata, as the original plants of the three Tibetan herbs, were collected and identified. The regions of ITS, matK, rbcL, rpoC1, trnL(UAA), psbA-trnH, atpB-rbcL, trnS (GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF(GAA) and nadl 2nd intron were amplified and sequenced. The ITS regions of S. hispidicalyx and S. dentata were cloned and sequenced, and the sequences were classified into different genotypes. All the sequences were analyzed and compared with those of closely related species. Our studies may provide reference for the genetic diversity analysis and molecular identification of the three Tibetan herbs.
Genetic Variation ; Gentiana ; classification ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; genetics ; Scrophularia ; classification ; genetics ; Swertia ; classification ; genetics ; Tibet

The alpine plant Gentiana robusta is an endemic species to the Sino-Himalayan subregion. Also, it is one of the original plants used as traditional Tibetan medicine Jie-Ji. We sequence the nuclear ribosomal internal transcribed spacer (ITS) regions, matK, rbcL, rpoC1, trnL (UAA), psbA-trnH, atpB-rbcL, trnS( GCU)-trnG(UCC), rpl20-rps12, trnL(UAA)-trnF( GAA) fragments of cp DNA in both G. robusta and such relative species as G. straminea, G. crassicaulis and G. waltonii. With Halenia elliptica as the outgroup, molecular systematic analysis reveals that G. robusta is a natural hybrid. G. straminea is the mother of hybrids, but the father is not very clear. In addition, the molecular markers for distinguishing G. robusta from the parental species or closely related species are identified, respectively. Our studies may provide valuable reference for the species identifications of medicinal plants with complex genetic backgrounds.
DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; Gentiana ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; genetics

Objective To study the mechanisms of Curcumin-induced apoptosis on human gastric cancer cell line SGC-7901.Methods SC,C-7901 cells were treated with various concentrations of Curcumin and the growth inhibition rates of it were accessed by MTT method.Apoptosis of gastric cancer cells were in- spected by flow cytometry.The expression of Fas and survivin in gastric cancer cells were evaluated by west- ern blot.Results Curcumin could effectively inhibit the growth of gastric cancer cells in dose-dependent and time-dependent manners,the sub-peak appeared and the apoptotic rate was increased.The expressions of Fas was higher in Western blot,meanwhile,the expressions of survivin was decreased.Conclusion Curcumin could significantly inhibit the growth and induce apoptosis of gastric cancer cells(SGC-7901),Curcumin could probably through up-regulating Fas and down-regulating surviving to induce apoptosis.

A strain was isolated from internal organ of died porcine about 8 weeks with purulent pneumonia,arthritis,pyogenic arthritis and endocarditis in April 2007.Objectives of the study are to confirm the genus of the strain,pathopoiesis,and drug sensitivity.The mainly study methods:the first,the strain was identified by the phenotype and the characteristics of the biochemistry,sequence 16S rDNA genes of the strain was analyzed by molecular biology technology,finally animal experiment and drug sensitivity testing were done.The results of the phenotype and the characteristics of the biochemistry showed that it is greatly similar to Actinomyces hyovaginalis,16S rRNA sequence analysis exhibited the homology achieved to 99.2% com-pared with group III strains of Actinomyces hyovaginalis,and the phyletic evolution analysis also indicated that it has mostly relationship with group III strains of Actinomyces hyovaginalis.Animal experiment dis-covered it has highly pathogenicity to Mus musculus albus;Drug sensitivity testing showed that it is hyper-sensitive to Erycin,Gentamicin and Amikacin.So,the result of the study confirmed that the strain is Actin-omyces hyovaginalis III with the pathogenicity.

Objective To detect the lymph node micrometastasis in resected pancreatic head carcinoma, to investigate the role of lymphatic micrometastasis in clinical staging and predicting prognosis of the pancreatic head carcinoma. Methods Pancreaticoduodenectomy with extended lymph nodes dissection were performed in 20 patients with pancreatic head carcinoma. All the lymph nodes were taken out by operating microscope method and metastasis was diagnosed by routine histological examination with hematoxylin and eosin staining, and the presence of lymph node micrometastasis was examined by immunohistochemisty. Results A total of 677 lymph nodes were found in the 20 eases, routine histological examination revealed metastasis occurred in 87 lymph nodes in 13 cases. Of the 590 negative lymph nodes by routine histological examination, 57 lymph nodes in 3 cases were diagnosed as having micrometastasis by immunohistochemisty. With the combination of routine histological examination and immunohistochemisty, the percent of patients with positive lymph nodes increased from 65% (13/20) to 80% (16/20), the detection rate of metastasis lymph node increased from 12.9% (87/677) to 21.3% (144/677) with significant difference (P <0.05). The detection of lymph node micrometastasis changed the staging of Ⅱ A to Ⅱ B in 3 patients. Tumor metastasis and recurrence rate of patients with lymph nodes micrometastasis within one year after operation was 75%, while it was 25% of patients without lymph nodes micrometastasis. Conclusions The detection of lymph node mierometastasis metastasis was helpful in the determination of clinical staging and predication of prognosis.

Objective To report the first case of cervical l ymphadenitis caused by Candida albicans in China.Methods A series of clinical,histopatholog ic.and mycologic studies were carried out in an 8year-old-boy with cervical lymphadenitis,who had no definite underlying disease.Candida albicans was isolated fromthe patient.The lymph node was investigated by me ans of culture,pathological and tra nsmission electron microscopic examination and cellular immunology detected by flow cytometer.Results There were several enlarged,warm,a nd fluctuant cervical lymph nodes which varied fr om 1cm?1cm to 4cm?4cm in size.One swo llen node was excised,and Candida albicans was found in the culture.The histopathologic changes of the cervical lym ph node were compatible with chronic granulomatous disease.PAS and methenamine silver(PAM)stain of a touch preparation of the bi opsy specimen revealed budding spores an d pseudohyphae.Lymphopenia with a p roportionate decrease of T-helper a nd T-suppressor cells was shown with flowcytometry.The patient received a combination therapy of surgery,flu conazole,thymosin and so on.After one month,t he patient's condition was improved.Many small lymph nodes disappeared,large lymph nodes became smaller.Tw o months later,only one large lymph n ode could be touched.Conclusion It is the first case of cervical lymphad enitis caused by Candida reported in China.Combination therapy of surgery,fluconazole and thymosin is effective.

Objective To establish an automatic warning program which accommodates to the test procedures in medical laboratory centerand and to put into operation.Methods Based on laboratory information management system (LIS),automatic warning program was established,which composed of maintenance of warning rules,feedback of vulnerabilities in rules,solutions of feedback targets and evaluation of ride-effectiveness.The results of performance were assessed after operation for six months in Guangzhou Kingmed Medical Laboratory Center.Results An automatic warning program containing 13 kinds of rule templates was successfully established.The multi-point warning program via asynchronous structure for six specialized testing procedures was realized.A total of 1 523 warning rules were included in the rule bank and 24 000 reports were verified on average each day.The average passing rate of reports was 70.2％ and the passing rate of single test was 83.9％.The approval time for reports was reduced by 10 to 40 minutes compared with that without using the automatic warning program.The efficiency for report approval was improved by 25 percent.Conclusion Automatic warning program may ensure the high quality of test reports,ease pressure on staff and improve work efficiency.

Objective The protective effect of hydrogen sulfide (H2 S) against myocardial ischemia/reperfusion ( IR) injury via anti-apoptotic signaling is well established , but the underlying mechanism remains unclear .This study was to investigate whether H 2 S could protect cardiomyocytes from endoplasmic reticulum stress ( ERS)-mediated apoptosis in hypoxia/reoxygenation ( HR) injury by regulating the expression of microRNA-455 ( miR-455 ) . Methods Cardiomyocytes from neonatal SD rats were primarily cultured and the model of HR injury was established .The cardiomyocytes were divided into a control group (normally cultured for 27 hours), an HR group (subjected to HR injury), and an H2S protection group (pretreated with the precursor of H2S NaHS at 40 μmol/L at 30 min before HR treatment followed by the same procedure as in the HR group ) .The cell viability was monitored by MTT , the release of lactate de-hydrogenase ( LDH) in the culture supernatant measured by full-automatic chemical analysis , and the apoptosis rate of the cardiomyo-cytes detected by flow cytometry .The mRNA and protein expressions of Grp 78 and caspase-12 were determined by real-time RT-PCR and Western bot .To verify whether miR-455 was involved in the ERS-mediated apoptosis of the cardiomyocytes , the cells were subjec-ted to HR after transfected with miR-455 mimic or anti-miR-455 oligonucleotide (AMO) for 24 hours, followed by detection of the ex-pressions of Grp78 and caspase-12. Results After HR injury, the H2 S protection group showed an enhanced viability of the cardio-myocytes in comparison with the control group ([67.02 ±6.90] vs [29.27 ±5.66] %), an decreased LDH release ([91.33 ± 10.63] vs [168.17 ±15.38] U/L), and a reduced rate of cell apoptosis ([13.98 ±1.90] vs [24.31 ±2.79] %).H2 S pretreat-ment significantly downregulated the mRNA and protein expressions of Grp 78 and caspase-12 (1.66 ±0.39 vs 2.56 ±0.34;1.75 ± 0.32 vs 2.54 ±0.48;2.01 ±0.45 vs 3.26 ±0.34;1.85 ±0.52 vs 3.21 ±0.84, P<0.05).The mRNA and protein expressions of Grp78 and caspase-12 were evidently increased after transfection with miR-455 mimic (3.56 ±0.37 vs 1.00 ±0.00;3.61 ±0.41 vs 1.00 ±0.00;2.87 ±0.38 vs 1.00 ±0.00;2.98 ±0.49 vs 1.00 ±0.00), but remarkably decreased after transfection with miR-455 AMO (0.62 ±0.16 vs 1.00 ±0.00;0.65 ±0.13 vs 1.00 ±0.00;0.54 ±0.13 vs 1.00 ±0.00;0.62 ±0.16 vs 1.00 ±0.00, P<0.05). Conclusion H2S could protect cardiomyocytes from HR injury by regulating the expression of miR-455 and reducing ERS-mediated cell apoptosis .

OBJECTIVETo establish an effective way for rapid identification of Monascus strains based on DNA molecular marker.

METHODA random amplified polymorphic DNA (RAPD) marker named F421 in genomic DNA of Monascus F strain was observed during a comparison of DNA fingerprints derived from 10 cultivated strains of Monascus. F421 was cloned and sequenced. Comparing the sequence of F421 (GenBank accession number EF063107) with other relative sequences in the GenBank databases, no distinct comparability was found. A pair of sequence characterized amplified region (SCAR) primers were designed based on the sequence of the cloned fragment and tested for the specific detection of Monascus F.

RESULTThe results of polymerase chain reaction showed that only a 421bp segment of Monascus F strain was amplified compared with other 9 cultivated strains of Monascus. And the acquired SCAR marker of strain F could be used as a specific DNA fingerprint to identify Monascus strain F within one day.

CONCLUSIONSCAR molecular marker technology is an effective new way to identify Monascus strains more rapidly. And also is an assistant tool to identify Monascus strains more accurately when disagreements come out using traditional classification. It could be applied widely to the protection of germ plasm resources, classification and identification distinguishing false strains of pharmaceutical fungi.

Molecular Sequence Data ; Monascus ; classification ; genetics ; Polymerase Chain Reaction ; methods ; Random Amplified Polymorphic DNA Technique

BACKGROUNDAsthma is a chronic inflammatory disease characterized by reversible bronchial constriction, pulmonary inflammation and airway remodeling. Current standard therapies for asthma provide symptomatic control, but fail to target the underlying disease pathology. Furthermore, no therapeutic agent is effective in preventing airway remodeling. A substantial amount of evidence suggests that statins have anti-inflammatory properties and immunomodulatory activity. In this study, we investigated the effect of rosuvastatin on airway inflammation and its inhibitory mechanism in mucus hypersecretion in a murine model of chronic asthma.

METHODSBALB/c mice were sensitized and challenged by ovalbumin to induce asthma. The recruitment of inflammatory cells into bronchoalveolar lavage fluid (BALF) and the lung tissues were measured by Diff-Quik staining and hematoxylin and eosin (H&E) staining. ELISA was used for measuring the levels of IL-4, IL-5, IL-13 and TNF-α in BALF. Periodic acid-Schiff (PAS) staining was used for mucus secretion. Gamma-aminobutyric acid type A receptor (GABAAR) β2 expression was measured by means of immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.

RESULTSRosuvastatin reduced the number of total inflammatory cells, lymphocytes, macrophages, neutrophils, and eosinophils recruited into BALF, the levels of IL-4, IL-5, IL-13 and TNF-α in BALF, along with the histological mucus index (HMI) and GABAAR β2 expression. Changes occurred in a dose-dependent manner.

CONCLUSIONSBased on its ability to reduce the inflammatory response and mucus hypersecretion by regulating GABAAR activity in a murine model of chronic asthma, rosuvastatin may be a useful therapeutic agent for treatment of asthma.

Animals ; Asthma ; drug therapy ; metabolism ; Chronic Disease ; Disease Models, Animal ; Female ; Fluorobenzenes ; pharmacology ; therapeutic use ; GABA-A Receptor Antagonists ; pharmacology ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Lung ; chemistry ; Mice ; Mice, Inbred BALB C ; Mucus ; secretion ; Pyrimidines ; pharmacology ; therapeutic use ; Receptors, GABA-A ; analysis ; Rosuvastatin Calcium ; Sulfonamides ; pharmacology ; therapeutic use