Genetic transformation is an effective method to improve breeding objective traits of orchids. However, there is little information about genetic transformation of Cymbidium sinensis. Rhizomes from shoot-tip culture of C. sinensis cv. 'Qijianbaimo' were used to establish a practical transformation protocol of C. sinensis. Pre-culture time, concentration and treating methods of acetosyringone, concentration of infection bacteria fluid (OD600), infection time, and co-culture time had significant effects on β-glucuronidase (GUS) transient expression rate of C. sinensis cv. 'Qijianbaimo' rhizome. The GUS transient expression rate of rhizome was the highest (11.67%) when rhizomes pre-cultured for 39 d were soaked in bacterium suspension (OD600 = 0.9) supplemented with 200 μmol/L acetosyringone for 35 min, followed by culturing on co-culture medium supplemented with 200 μmol/L acetosyringone for 7 d. Under this transformation conditions, 3 transgenic plantlets, confirmed by GUS histochemical assay and PCR, were obtained from 400 regenerated plantlets, and the genetic transformation rate was 0.75%. This proved that it was feasible to create new cultivars by the use of Agrobacterium-mediated genetic transformation in C. sinense.
Agrobacterium ; Coculture Techniques ; Genetic Engineering ; Glucuronidase ; Orchidaceae ; genetics ; Plants, Genetically Modified ; genetics ; Polymerase Chain Reaction ; Transformation, Genetic

Diversified teaching methods including traditional teaching,problem based learning,interactive teaching and team cooperation teaching were adopted in the process of teaching.Problem based learning can stimulate students' learning interests; interactive teaching can improve the students' hands-on ability and team cooperation teaching can cultivate students' cooperation spirits.Diversified teaching mode made different teaching methods integrated and complemented each other,forming a special teaching mode for the course of functional experiment.

ObjectiveToinvestigatetheroleofimmuneinflammatoryreactionintheformationof intracranial aneurysm. Methods The intracranial aneurysms in 40 patients of craniotomy ( intracranial aneurysm group) and the vascular specimens in 20 craniotomy patients w ith traumatic brain injury (control group) w ere col ected. Fluorescence quantitative polymerase chain reaction w as used to detect the expression of interleukin (IL)-17 receptor in the arterial w al . Flow cytometry w as used to detect the Th-17 cel s in peripheral blood. Enzyme-linked immunosorbent assay w as used to measure the levels of IL-17, IL-6 in the arterial w al and tumor necrosis factor-α( TNF-α) in peripheral blood. Results There w ere no significant differences in the age (62.6 ±8.7 years vs.61.4 ±7.9 years;t=0.342;P=0.681), proportions of male (60.0%vs.65.0%; χ2 =0.246, P=0.434), hypertension ( 12.5%vs.10.0%; χ2 =0.315, P=0.492), diabetes (75.0%vs.10.0%; χ2 =0.284, P=0.482), and smoking (35.5%vs.30.0%; χ2 =0.224, P=0.413) betw een the intracranial aneurysms group and the control group. The expression of IL -17 receptor in the arterial w al (0.106 ±0.032 vs.0.264 ±0.071; t=5.115, P=0.001) and the proportion of Th17 cels in peripheral blood (2.75%±0.53%vs.7.18%±1.54%; t=8.436, P<0.001) and IL-17 level ( 7.32 ±1.82 μg/L vs.22.64 ±4.51 μg/L; t= 8.357, P< 0.001 ) in the control group w ere significantly low er than those in the intracranial aneurysm group. The levels of IL-6 (1.15 ±0.24 μg/L vs. 19.64 ±4.16 μg/L; t=9.527, P<0.001) and TNF-α(1.43 ±0.31 μg/L vs.26.17 ±4.32 μg/L; t=9.816, P<0.001) in the arterial wal in the control group were significantly lower than those in the intracranial aneurysm group. Conclusions The expression of IL-17 receptor in the arterial w al , the proportion of the Th17 cels and IL-17 level in peripheral blood were increased in patients with intracranial aneurysms. Immune inflammation may be involved in the formation of intracranial aneurysm.

Objective: To observe the clinical efficacy of acupuncture plus navel acupuncture for patients with urinary retention after radical hysterectomy for cervical cancer. Methods: A total of 64 patients with urinary retention after radical hysterectomy for cervical cancer was divided into a navel acupuncture group (22 cases), an acupuncture group (18 cases) and an acupuncture plus navel acupuncture group (24 cases). All three groups received bladder function training and neuromuscular electrical stimulation. In addition, navel points were combined in the navel acupuncture group. Electroacupuncture was conducted to Qihai (CV 6), Zhongji (CV 3), Dahe (KI 12), Shuidao (ST 28), Ciliao (BL 32) and Huiyang (BL 35) in the acupuncture group. The acupuncture plus navel acupuncture group received both treatments. The catheter was removed after 3 d of treatment. Spontaneous urination, residual urine volume, urinary catheter dependence and recurrence after 3 d, 6 d and 9 d of treatment in each group were observed, respectively. Results: In the acupuncture plus navel acupuncture group, the markedly effective rates after 3 d, 6 d and 9 d of treatment were significantly higher than those in the navel acupuncture group and the acupuncture group; the urinary catheter dependence was lower than that of the other two groups, and the differences were statistically significant (P<0.05, P<0.01); the spontaneous urination time was shorter than that of the navel acupuncture group and the acupuncture group, and the differences were statistically significant (P<0.05, P<0.01); the residual urine volume was significantly less than that of the navel acupuncture group and the acupuncture group, and the differences were statistically significant (both P<0.01). After the catheter was removed, recurrence was observed from the next day after spontaneous urination was resumed. There were 2 cases of recurrence in the navel acupuncture group, 2 cases in the acupuncture group and 1 case in the acupuncture plus navel acupuncture group. The recurrence rate of the acupuncture plus navel acupuncture group was significantly lower than that of the navel acupuncture group and the acupuncture group (both P<0.01). Conclusion: Acupuncture plus navel acupuncture has satisfactory efficacy for urinary retention after radical hysterectomy for cervical cancer. It can significantly shorten the urinary retention time, reduce the patient's dependence on urinary catheter, and reduce the residual urine volume.

Adolescent ; Adult ; Agranulocytosis ; microbiology ; Anti-Bacterial Agents ; adverse effects ; therapeutic use ; Daptomycin ; adverse effects ; therapeutic use ; Female ; Gram-Positive Bacterial Infections ; drug therapy ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVEThis study was aimed to characterize the Helicobacter pylori strains isolated from different geographic regions in China and different ethnic groups in Yunnan province in terms of cagA, iceA, vacA and HP0519 genes which were proposed to be related to the pathogenesis.

METHODS150 Helicobacter pylori strains were collected from Yunnan province, Fujian province and Beijing. Chromosome DNA was extracted and polymerase chain reaction (PCR) was carried out to determine the 3' region of cagA, iceA, vacA and HP0519 status with specific primers. PCR results were analyzed statistically according to their isolated original and clinical outcomes.

RESULTSFor cagA 3' region, 93% (139/150) of the Chinese Helicobacter pylori strains belonged to East Asian type according to the specific primer of TF/JR. Among the 150 strains, 75% (113/150) belonged to iceA1, and 19% (29/150) to iceA2. The dissemination of iceA was not associated with any of the geographic regions, different ethnic groups or different clinical outcomes. 96% (144/150) of the vacA s region belonged to s1. In the vacA middle region, m2, m1b, m1b-m2 were 57% (85/150), 27% (41/150) and 11% (16/150) respectively. However, m1a was only observed in two strains from Fujian. Neither vacA s1 nor m2 showed significant difference between Yunnan, Fujian and Beijing. However, the distribution of mlb-m2 in Yunnan was higher than that in Fujian and Beijing. In Yunnan province, the distribution of vacA s1 was not associated with different ethnic groups but m2 from Bai group was less than other two ethnic groups. The ratio of m1b in Bai group was higher than that in other groups. Both vacA' s region and m region alleles had no significant relationship with the clinical outcomes. With the 15 bp and 24 bp DNA insertion and deletion primers test, 93% (140/150) of the strains were positive. The distributions of the 15 bp and 24 bp DNA insertion or deletion were different according to the different ethnic groups.

CONCLUSIONBy JF/TR primer, 93% of the Chinese strains cagA's 3' region belonged to East Asian type. Most of the Chinese strains vacA's allele was s1. The distribution of vacA s1 had no relationship with the clinical outcome of the isolates. From different geographic regions and ethnic groups, the distribution of vacA m region allele was different. 93% of the Chinese strains HP0519 genes had 24 bp or 15 bp insertion or deletion character. The biological meaning of the polymorphism of HP0519 needs advanced investigation.

China ; Genes, Bacterial ; genetics ; Helicobacter Infections ; ethnology ; genetics ; Helicobacter pylori ; classification ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction

OBJECTIVETo investigate the prevalence of chronic obstructive pulmonary disease (COPD) and its risk factors in population over 40 years old in northern part of Guangdong province.

METHODSUsing uniform scheme, procedures and questionnaire, a cluster-randomized-sampling survey for the population aged over 40 years in a rural area of Shaoguan in the northern part of Guangdong province was performed. Spirometry was performed for every participant, followed by a bronchodilatation test when bronchial obstruction was present.

RESULTSThere were 1468 cases with complete data from 1498 people aged >or= 40 years including 640 males, 828 females with an average age of 54.3 years old. The total prevalence of COPD was 12.0%. The prevalence of COPD in males was significantly higher than that in females (18.3% vs. 7.1%, P < 0.01). Only 80.7% of the patients with COPD presented one or more symptoms as cough, phlegm, or dyspnoea. Underdiagnosis of COPD would be quite serious. Only 26.1% of the cases was previously diagnosed to have chronic bronchitis, emphysema, or COPD. Smoking was an important risk factor to COPD and 78.4% of the patients with COPD were smokers. However, relation of biomass and COPD called for further investigation.

CONCLUSIONPrevalence of COPD was much higher than expected in the northern part of Guangdong while smoking was an most important risk factor of COPD. Lung function test seemed to be of great importance to COPD diagnosis, especially in the earlier period of COPD.

Adult ; China ; epidemiology ; Female ; Humans ; Male ; Mass Screening ; Middle Aged ; Prevalence ; Pulmonary Disease, Chronic Obstructive ; epidemiology ; Risk Factors ; Sex Factors ; Smoking ; adverse effects ; Surveys and Questionnaires

OBJECTIVETo explore a flow cytometry (FCM)-based method for discriminating aneugen- or clastogen-induced micronuclei.

METHODSCells were stained with anti-CD71-FITC and PI, and the PI fluorescent signal intensity of micronucleated reticulocyte (MN-RET) in the peripheral blood of NIH mouse treated with COL or CP was detected by flow cytometry.

RESULTSThe ratio of the median of the intensity of MN-RET fluorescent signals to that of nucleated cell was low in the cyclophosphamide treated mouse, while the median was high in the colchicine treated mouse.

CONCLUSIONThe flow cytometry-based micronucleus assay can be used to discriminate primarily smaller MN induced by the clastogen exposure from the larger MN induced by an aneugen.

Animals ; Colchicine ; toxicity ; Cyclophosphamide ; toxicity ; Flow Cytometry ; methods ; Male ; Mice ; Micronuclei, Chromosome-Defective ; Mutagens ; toxicity ; Reticulocytes ; drug effects ; ultrastructure

OBJECTIVETo evaluate the ancillary diagnostic value of IgH gene rearrangements in those B-cell lymphoproliferative disorder cases whom are difficult in making a final diagnosis.

METHODSIgH gene clonal rearrangements were retrospectively analyzed in a total of 77 diagnostically difficult B-cell lympho-proliferative patients. Standardized BIOMED-2 system IgH gene clonality assay kit targeting FR1, FR2, FR3 was used, followed by heteroduplex-polyacrylamide gel electrophoresis (PAGE) and silver nitrate staining.

RESULTSThe final diagnoses of the 77 cases were: 12 cases of reactive lymphoid hyperplasia, 20 cases of atypical lymphoid hyperplasia or suspicious lymphoma, and 45 cases of B-cell lymphoma. Detection rates of at least one positive reaction were 2/12, 11/20 (55%), 36/45 (80%) in the three groups, respectively. In B-cell lymphomas, the clonality detection rate of FR1, FR2 and FR3 was 60% (27/45), 60% (27/45) and 56% (25/45), respectively. The type distribution were: 20 marginal zone lymphomas, including 18 extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue, 7 diffuse large B-cell lymphomas, 7 follicular lymphomas, 1 mantle-cell lymphoma, 1 Burkitt's lymphoma, 4 plasma cell neoplasms and 5 unclassified B-cell lymphomas. Rearrangements of FR1, FR2 or FR3 were not detected in 9 (20%) of the B cell lymphoma cases, nevertheless, one of them had developed liver lesion later, and was confirmed finally to be B cell lymphoma. Fourteen patients of reactive lymphoid hyperplasia with positive IgH gene clonal rearrangements, and atypical lymphoid hyperplasia had follow-up history available. Four of them were diagnosed as lymphoid malignancies upon further biopsy, and in three of them, clonal IgH gene rearrangements were detected.

CONCLUSIONSB-cell lymphoproliferative disorder requiring a detection of clonal IgH gene rearrangement for making a final diagnosis. Combined detections of three IgH FR1, FR2 and FR3 rearrangements provide important ancillary diagnostic value in confirming suspected B-cell lympho-proliferative disorders. It is important to take an additional biopsy or to follow-up those patients who that have a detectable IgH gene clonal rearrangement but without apparent morphological evidence of lymphoma. For cases with a negative IgH gene rearrangements, it might be necessary to perform clonality analysis for other forms of gene rearrangements including IgH or IgK and IgL in order to further improve the detection sensitivity.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Humans ; Lymphoma, B-Cell ; diagnosis ; genetics ; pathology ; Lymphoma, B-Cell, Marginal Zone ; diagnosis ; genetics ; pathology ; Lymphoma, Follicular ; diagnosis ; genetics ; pathology ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; genetics ; pathology ; Lymphoproliferative Disorders ; diagnosis ; genetics ; pathology ; Male ; Middle Aged ; Neoplasms, Plasma Cell ; diagnosis ; genetics ; pathology ; Pseudolymphoma ; diagnosis ; genetics ; pathology ; Retrospective Studies ; Young Adult

OBJECTIVETo evaluate the prevalence of Helicobacter pylori (H.pylori) resistance to metronidazole among three populations in Yunnan.

METHODSSusceptibilities to metronidazole among 109 H. pylori strains (33 H. pylori strains from Han, 31 H. pylori strains from Bai and 45 H. pylori strains from Naxi ethnic populations) were tested by Epsilometer test (E-test).

RESULTSIn 109 H. pylori strains, the overall metronidazole resistance rate was 67.89%. There were no significant difference in the metronidazole resistant rates of H. pylori among Han, Bai, Naxi populations Yunnan in terms of the distribution on age and upper gastroduodenal diseases. In the facet of gender, metronidazole resistant rate of H. pylori was significantly lower in Han males than in females (chi2=5.304, P=0.027), but not seen in the Bai or Naxi peoples.

CONCLUSIONMetronidazole resistance rate of H. pyloriin Yunnan was high, but no significant difference was found among Han, Bai, Naxi peoples in the province.

Adult ; China ; ethnology ; Chronic Disease ; Drug Resistance, Bacterial ; Female ; Gastritis ; microbiology ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; drug effects ; Humans ; Male ; Metronidazole ; pharmacology ; therapeutic use ; Middle Aged ; Peptic Ulcer ; microbiology