In plant genetic engineering, selectable marker is needed to distinguish transformant. As the commercialization of transgenic plants, people are more and more paying close attention to their safety, among which mainly refers to the safety of selectable marker. In order to increase the safety of transgenic plants, biologists began to search for biosafe selectable marker.Current research progress of taking key enzyme in metabolic pathways were overviewed,which include glycometabolism, amino acid metabolism, hormone metabolism ,nucleotide metabolism and protein metabolism et cetera, as selectable marker in transgenic plants.

The rodipicolinate synthase gene was cloned from Nicotiana tabacum L.cv,which encodes the key enzyme in the synthesis of lysine,and then a vitro molecular reform reliefing feedback suppression was made. Some resistant seedlings were acquired using the mutated gene as selectable marker and analog of lysine as selectable reagent,and the detection results were positive by means of PCR and Real-Time PCR,however the phenotype is somewhat abnormal.

10-15 years were 7 cases(6.3%),they were randomly divided into treatment group and contol group.The treatment group with 60 patients were given INF-? and polyresistin,age 3 years,given IFN-? 200 000 U per day;and polyresistin 5 mg per time,3 times every day,the course of therapy were 5 days.The control group was given traditional Chinese medicine at the same time.Results After therapy,in treatment group,53 cases(88.3%) of children′s cooling time were lower than 5 days,47 cases(78.3%) of children′s rash subsided time were lower than 6 days,48 cases(75%) of children′s Catarrhal symptoms disappeared time were lower than 5 days,and the rate of complications′ occurrence were 15 cases(18.3%).In control group,28 cases(53.8%) of children′s cooling time were lower than 5 days,14 cases(26.9%) of children′s rash subsided time were lower than 6 days,28 cases(53.8%) of children′s Catarrhal symptoms disappeared time were lower than 5 days,and the rate of complications′ occurrence were 37 cases(71.5%),there were significant differences between both groups(Pa

Objective To explore whether the transcription factor NF-κB takes part in baicalin-induced differentiation of bone marrow stromal cells (MSCs) into neurons. Methods MSCs from adult rats were induced by baicalin in the serum-free medium for 6 hs. The un-treated cells, as the control group, were only induced in the serum-free medium without baicalin. The expression of neuronal specific markers was evaluated by indirect immunofluorescence cytochemistry staining. The activity of NF-κB was measured by the presence of the NF-κB subunit RelA (p65) translocated into the nucleus in the same way. Results After the induction by baicalin, MSCs displayed neuronal morphologies, such as pyramidal cell bodies and extensive networks of processes, and the expression of neuron-specific markers was detectable 6 ds after the induction. Neuronal specific marker proteins did not express in the control group. Six days after the induction, the P65 positive rate in the cytoplasm of the control group decreased to (18.4±3.0)%, while in the baicalin group, the P65 positive rate in the cytoplasm was (84.8±3.0)%. Conclusions Baicalin may inhibit the activation of NF-κB, which may act in the differentiation of MSCs into neurons.

Objective To explore the protocol that induced marrow mesenchymal stem cells (MSCs) differentiating into islet-secreting cells in vitro and to provide new clues for the sources of islet transplantation.Methods Using a defined culture medium and technique for transdifferentiation, MSCs from adult SD rats were guided into specific insulin-secreting cells. The expressions of nestin and islet-specific hormones and proteins, such as insulin, glucagon, somatostatin and pancreatic duodenal homeobox 1 (Pdx-1) were analyzed by indirect immunofluorescence cytochemistry staining before and after induction. The expressions of pancreatic islet cell differentiation-related transcripts, such as nestin, insulin 1, glucose transporter 2 (GLUT 2), Isl-1, Pdx-1, Pax-4 and Pax-6 were detected by reverse transcription-PCR (RT-PCR). In addition, the quantity of insulin secretion was examined using radioimmunoassay. Results Five hours after induction, (44.6±7.3)% of differentiated MSCs expressed nestin and it increased to (61.8±8.4)% 24 hs after induction, but the expression of nestin almost disappeared at day 14. In the meantime, islet-like cellular clusters appeared after day 14 and became more apparent by day 28. Differentiated cells were found to be immunoreactive to insulin, glucagon, somatostatin and Pdx-1, and expressed insulin 1, GLUT 2, GK, Isl-1, PDX-1, Pax-4, Pax-6 mRNA. In addition, the results of cumulative quantities of insulin of 24 hs and the stimulation index showed that differentiated cells were able to produce insulin at higher levels, and displayed glucose-dependent insulin release in vitro. Conclusions Adult rat MSCs can be differentiated into insulin-secreting cells in vitro. This approach might lead to widespread cell replacement therapy for Type 1 diabetes.

BACKGROUND: As a primary clinical predictor of fracture risk, bone mineral density (BMD) is partly genetically determined. Osteoprotegerin (OPG) is one important candidate gene in the pathogenesis of osteoporosis. OBJECTIVE: To investigate the association between T245G polymorphisms in the OPG gene and BMD. METHODS: A total of 281 elderly men and postmenopausal women, 182 males and 99 females, who received routine examinations at Peking University People's Hospital between September 2008 and April 2010 were included in this study. T245G polymorphisms in the OPG gene was detected by polymerase chain reaction-restriction fragment length polymorphism together with DNA sequencing. The BMD of the lumbar spine, Ward's triangle, and forearrm was determined by dual energy X-ray absorptiometry. Clinical variables and biochemical measurements were collected simultaneously. The association between T245G polymorphisms and each detection index was analyzed using analysis of variance. RESULTS AND CONCLUSION: The distribution of T245G genotype (alleles T, G) had no difference in elderly men or postmenopausal women (P > 0.05). The GG genotype and TG genotype had higher lumbar spine BMD and TT genotype had lower lumbar spine BMD (P < 0.05). There was no difference in BMD of the Ward's triangle or forearm among different genotypes (P > 0.05). Association between T245G polymorphism and BMD was not found in postmenopausal women. These findings indicate that OPG gene is related to lumbar spine BMD in elderly men.

In this paper 3 different media (A,for yeast cultivation ; B, for laccase producing; D, for white rot fungi cultivation) were compared in enriching and screening decolorizing fungi culture using Reactive Black 5 (RB5) and Reactive Red (M-3BE) from the following three points: decolorization effects, abilities of producing enzymes and diversity of microbial community. 11 groups of fungi with obvious decolorization effects were obtained after enrichment for near one month. Among them, 6 groups came from medium D, the other two 3 groups from medium A and B, respectively. However, the 3 groups from medium A exhibited the highest microbial diversity and best decolorization results with 99.53% and 97.42% color removal rate of Reactive Red M-3BE and Acid Red. From them, 16 strains of fungi were isolated and primarily identified as Saprolegniaceae, Eurotiaceae (Monascus went), Erysiphaceae and Physodermataceae. Fungi groups from medium B and D exhibited a bit lower color removal rate of various dyes and only 3 and 2 isolates primarily classified as Saccharomycetaceae and Eurotiaceae (Penicillium) were obtained from them. Fungi cultures in medium A and B could produce lignin peroxidase, and those in medium D could be detected higher activity of laccase. All the fungal cultures exhibited very weak activity of manganese dependant peroxidase.

OBJECTIVETo observe the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer, and study its anti-tumor mechanism.

METHODIn vitro, MTT assay and scratch assay were adopted to detect the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer cells. The cell autophagy was detected by the acridine orange staining. The gap-junction intercellular communication (GJIC) was investigated by the fluorescent yellow transfer. The expression of aquaporin 1 (AQP1) was analyzed by the Western blotting. In vivo, the subcutaneous implant model and the experimental pulmonary metastasis model of Lewis lung cancer in mice were established to evaluate the anti-tumor and anti-metastasis effects of alcohol extract from Pharbitidis Semen. The serum carcinoembryonic antigen (CEA) and beta2 microglobulin (beta2-MG) of mice bearing Lewis lung cancer were detected by the electrochemiluminesence immunoassay. The expressions of lung AQP1 and Connexin 43 (Cx43) were examined by the immunohistochemical method.

RESULTIn vitro, alcohol extracts from Pharbitidis Semen inhibited the cell proliferation in a dose-dependent matter, significantly prevented the cell migration, down-regulated AQP1 proteins of cells, promoted GJIC, and decreased the serum-free autophagy of tumor cells. In vivo, compared with untreated model mice, alcohol extracts from Pharbitidis Semen inhibited the tumor growth in a dose-dependent matter, prevented the tumor metastasis and prolonged the life span of mice bearing Lewis lung cancer, while decreasing serum CEA and beta2-MG of mice bearing Lewis lung cancer, enhancing the immumohistochemical staining intensity of Cx43 and weakening aquaporins AQP1 positive intensity.

CONCLUSIONAlcohol extracts from Pharbitidis Semen could prevent the proliferation and metastasis in Lewis lung cancer cells. Its mechanism may be related to the promotion of GJIC and the down-regulation of AQP1.

Animals ; Antineoplastic Agents ; administration & dosage ; Aquaporin 1 ; genetics ; metabolism ; Carcinoma, Lewis Lung ; drug therapy ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Connexin 43 ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Ipomoea ; chemistry ; Lung Neoplasms ; drug therapy ; genetics ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Metastasis ; Seeds ; chemistry

Objective In order to investigate the relationship between hearing loss and diabetic peripheral neuropathy( DPN ) via comparing the pure tone thresholds and vibration perception threshold(VPT) in type 2 diabetic patients and control person without diabetes. Methods 173 subjects including 138 type 2 diabetic patients(DM)and 35 non-diabetes controls were examined for VPT and hearing threshold. Nerve conducting velocity ( NCV )including sensory nerve conducting velocity( SCV )and motorial nerve conducting velocity( MCV )of diabetic patients were determined. The participants were divided into three groups: control group (n = 35 ), DM group without peripheral neuropathy( non-PN group, n = 74 ), and DM group complicated with peripheral neuropathy (PN group,n = 64 ). The clinical characteristics, biochemical parameters , the incidence of sensorineural hearing loss ( SNHL),pure tone threshold, and VPT were compared among three groups. At last, the relationship between hearing thresholds and NCV were analyzed. Results The incidence of hearing impairment of sensorineural type was 29.69% in PN group, which was significantly higher than that of non-PN group( 17.57% )and control group( 17. 14% ). There was significant differences in age, duration of diabetes, glycolated hemoglobin (HbA1c), glycolated serum albumin ( GA), Fasting blood glucose( FPG), 2h postprandial blood glucose( PPG), VPT, and hearing threshold among the three groups( all P＜0. 05 ). The value of hearing threshold increased significantly( all P＜0. 05 ) in 3 VPT subgroups with VPT≤ 15 V, VPT 16-25 V, and VPT ＞25 V. The Spearman correlation analysis showed median NCV was negatively correlated with hearing threshold on 1.00, 2.00, 4. 00, and 8. 00 kHz ( All P ＜ 0. 05 ). The logistic regression analysis indicated that the age( regression coefficient =0. 088, P＜0. 01 ) was the independent risk factor of SNHL, median nerve MCV ( regression coefficient = -0. 135, P = 0. 046 ) was the important influencing factor of SNHL. Conclusion Diabetic patients are more likely to suffer from impaired middle-frequency and high-frequency hearing, DPN in patients is often complicated with hearing impairment. Age and median nerve MCV were major risk factors of SNHL in diabetic patients.

Adult ; Female ; Humans ; Intestinal Obstruction ; etiology ; Sjogren's Syndrome ; complications ; drug therapy