Female ; Humans ; Infant ; Leukocyte Elastase ; isolation & purification ; Neutropenia ; congenital ; enzymology

This study is to observe the protection of gross saponins of Tribulus terrestris (GSTT) on cardiocytes impaired by adriamycin (ADR) and approach its mechanism of action. Cardiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, model (ADR 2 mg x L(-1)) group, and GSTT (100, 30, and 10 mg x L(-1)) groups. MTT colorimetric method was deployed to detect cardiocyte survival rate, activities of CK, LDH, AST, SOD, MDA and NO were detected, and apoptosis was detected with flow cytometry. Effect of GSTT on caspase-3 was detected with Western blotting. Compared with control group, contents of CK, LDH, AST, MDA and NO were increased, and activity of SOD was reduced (P < 0.05, P < 0.01, P < 0.001) by ADR. Numbers of survival cells were increased (P < 0.05, P < 0.001), contents of CK, LDH, AST, MDA and NO were decreased, and activity of SOD was increased (P < 0.05, P < 0.01, P < 0.001) by GSTT (100 and 30 mg x L(-1)). Apoptosis of cardiocytes and concentration of caspase-3 can be reduced by GSTT (100 and 30 mg x L(-1)). GSTT can protect cardiocytes impaired by ADR, which are possible involved with its effect of resisting oxygen free radical.

This study is to observe the protection of gross saponins of Tribulus terrestris (GSTT) on cardiocytes impaired by adriamycin (ADR) and approach its mechanism of action. Cardiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, model (ADR 2 mg?L-1) group, and GSTT (100, 30, and 10 mg?L-1) groups. MTT colorimetric method was deployed to detect cardiocyte survival rate, activities of CK, LDH, AST, SOD, MDA and NO were detected, and apoptosis was detected with flow cytometry. Effect of GSTT on caspase-3 was detected with Western blotting. Compared with control group, contents of CK, LDH, AST, MDA and NO were increased, and activity of SOD was reduced (P

In this paper, the chloroplast psbK-psbI intergenic spacers of 18 species of Dendrobium and their adulterants were amplified and sequenced, and then the sequence characteristics were analyzed. The sequence lengths of chloroplast psbK-psbI regions of Dendrobium ranged from 474 to 513 bp and the GC contents were 25.4%-27.6%. The variable sites were 71 while the informative sites were 46. The inter-specific genetic distances calculated by Kimura 2-parameter (K2P) of Dendrobium were 0.006 1-0.058 1, with an average of 0.028 4. The K2P genetic distances between Dendrobium species and Bulbophyllum odoratissimum were 0.093 2-0.120 4. The NJ tree showed that the Dendrobium species can be easily differentiated from each other and 6 samples of the inspected Dendrobium species were identified successfully through sequencing the psbK-psbI intergenic spacer. Therefore, the chloroplast psbK-psbI intergenic spacer can be used as a candidate marker to identify Dendrobium species and its adulterants.
Chloroplasts ; DNA, Chloroplast ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Plants, Medicinal ; classification ; genetics

Cooperation hydrogen production was carried out using Rhodopseudomonas sp. DT and Enterobacter aerogenes. The effects of the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes, culture temperature, and carbon source on the cooperation hydrogen production were investigated. The results suggested that cooperation hydrogen production rate was highly affected by the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes. The mixed bacteria of Rhodopseudomonas sp. DT and E. aerogenes with 1:1 initial ratio benefited to the cooperation hydrogen production, which led the hydrogen production rate and duration of gas production to 3.1 mol H2/mol glucose and 81 h, respectively. The pH dynamics analysis of culture medium further discovered that the pH of the mixed bacteria with 1:1 initial ratio changed from 6 to 7 smaller than other conditions, which was probably fitted to produce hydrogen. Furthermore, the mixed bacteria with 1:1 initial ratio had the higher hydrogen production efficiency at temperatures of 28?C and 37?C than at 20?C, and without any hydrogen production at temperature of 50?C. The carbon sources of glucose, succinate acid, malic acid could be used to produce hydrogen by the mixed bacteria. Even the soluble starch, unused by Rhodopseudomonas sp. DT, was also decomposed by the mixed bacteria to produce hydrogen with the conversion efficiency of 8.22%. The glucose was the optimal carbon resource, and the conversion efficiency could reach to 36.11%. The results, further, implied that the cooperation hydrogen production could enlarge the use of the carbon sources.

To purify the recombinant human BMP-6 protein and to establish its in vitro bioassay method. The cDNA encoding the mature peptide of hBMP-6 protein was amplified by reverse transcription-polymerase chain reaction (RT-PCR), using human placental mRNA as template, and subcloned into the high-expression vector pET-15b under the control of T7 lac promoter. The resulting construct, pET-BMP6, was then transformed into an Escherichia coli strain BL21 (DE3) for the production of recombinant hBMP-6 protein (rhBMP-6). After 4 hours of induction by isopropyl-beta-D-thiogalactoside (IPTG), rhBMP-6 (approximately 15kD) was expressed and formed inclusion bodies, contributing up to 10% of the total bacterial protein. The inclusion bodies were isolated and redissolved in 8mol/L urea, and the denatured rhBMP-6 was purified to 95% purity by CM-Cellulose 32 ion exchange chromatography (IEC). The osteoinductivity of rhBMP-6 was measured by the expression of some of the osteoblast differentiation marker genes in rhBMP-6-treated C3H10T1/2 cells as reflected by determinations of alkaline phosphatase (ALPase) activity and semi-quantitative RT-PCR. At the end of the purification process, about 80% of rhBMP-6 formed disulphide-linked homodimers after refolding during renaturation. The apparent size of the protein was 30kD on non-reducing SDS-PAGE, similar to that of the native form of hBMP-6. The enzyme assays showed that the ALPase activity was increased in a dose-dependent manner with the treatment of rhBMP-6. After the addition of 300ng/mL of rhBMP-6, the ALPase activity of C3H10T1/2 cells increased significantly. The activity of rhBMP-6 used was comparable to about 70% of that of the standard hBMP-6 derived from eukaryotic cells. RNA extraction data also showed rhBMP-6 stimulated expression of osteoblast marker genes, including type I collagen, osterix, and osteocalcin in a time-dependent manner. After 5 days of treatment, their level of expression was increased to 3 times that of controls. Bone morphogenetic protein (BMP)-6, a member of the 60A subgroup of the bone morphogenetic protein (BMPs) family, plays a pivotal role in bone formation. Previous evidence showed that BMP-6 is selectively up-regulated by estrogen, suggesting its potential role in the treatment of osteoporotic fractures, especially for menopausal osteoporosis. Our present study demonstrates that the recombinant hBMP-6 produced in Escherichia coli is able to induce pre-osteoblastic cells to differentiate into osteoblasts in vitro, and analysis of mRNA expression of type I collagen, osterix, and osteocalcin can be also a method for measuring the osteoinductivity of BMP. This provides the basis for further studies on ectopic bone formation in the body and for the development of auxiliary drugs for the treatment of osteoporotic fractures.
Alkaline Phosphatase ; metabolism ; Animals ; Biological Assay ; methods ; Bone Morphogenetic Protein 6 ; genetics ; isolation & purification ; metabolism ; pharmacology ; Cell Line ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Enzyme Activation ; drug effects ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; drug effects ; genetics ; Humans ; Isopropyl Thiogalactoside ; pharmacology ; Mice ; Plasmids ; Reverse Transcriptase Polymerase Chain Reaction

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; genetics ; Female ; Genes, erbB-1 ; genetics ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Nucleic Acid Amplification Techniques ; methods ; Receptor, Epidermal Growth Factor ; genetics

Objective To observe the therapeutic effect of Lai's acupuncture therapy combined with Chinese herbal decoction for the treatment of infertility with ovulatory dysfunction, and to summarize the experience of Professor Lai Xin-Sheng in treating infertility. Methods Ninety-six infertility patients with ovulatory dysfunction were randomized into treatment group and control group, 48 cases in each group. The treatment group was treated with Lai's Tongyuan acupuncture therapy for regulating Governor Vessel to tranquilize mind, inducing meridian Qi to original Qi, and was also given oral use of Chinese herbal decoction of Lai's Yiyun Recipe for promoting gestation. The control group was treated with Chinese herbal decoction alone. The treatment began exactly after the completion of menstruation period and lasted 15 days for one treatment course, covering 3 courses. And then the patients of the two groups were followed-up for 3 months. Therapeutic effect and pregnancy rate were evaluated after treatment, serum levels of estradiol and progesterone were examined, and abdominal ultrasonography was used for monitoring the maximum diameter of the dominant follicles and endometrial thickness before and after treatment. Results (1) Three cases from the treatment group and 5 cases from the control group were dropped out, and the final case number in the treatment group and control group was 45, 43 respectively. (2) The total efficiency of the treatment group was 88.9％, and that of the control group was 81.4％. The effect of the treatment group was better than that of the control group (P < 0.05). (3) The pregnancy rate of the treatment group was 73.3％, higher than that of the control group (51.2％), the difference being significant (P < 0.05). (4) After treatment, the maximum diameter of the dominant follicles and endometrial thickness in the two groups were obviously improved(P < 0.05 compared with those before treatment), and the improvement in the treatment group was superior to that in the control group (P < 0.05). (5) After treatment, the serum levels of estradiol and progesterone in the two groups were obviously improved(P < 0.05 compared with those before treatment) , and the improvement in the treatment group was superior to that in the control group(P < 0.05). Conclusion The therapeutic effect of Lai's acupuncture therapy combined with Chinese herbal decoction is superior to that of Chinese herbal decoction alone, showing an obvious synergistic effect by shortening the course of treatment and enhancing pregnancy rate.

Objective To explore the expression of chemerin and chemerin receptor ( chemokine-like receptor 1, CMKLR1) during different periods of non-alcoholic fatty liver disease ( NAFLD) rat model induced by methionine- and choline-deficient ( MCD) diet. Methods Thirty-six Wistar rats were divided into control group and MCD group in random. After one week quarantine and acclimation period, these two groups were fed either normal chow or MCD diet. The animals were respectively sacrificed at the first week, the forth week, and the tenth week. The levels of alanine transaminase (ALT), blood lipid profile, liver function, and the content of triglyceride in liver were detected. HE staining was done to observe the morphologic change of liver. The mRNA expression changes of chemerin and CMKLR1 in liver were measured using real-time PCR, and the change in chemerin mRNA level was further confirmed in liver by Northern blot. Finally, the concentration of chemerin in serum was measured by Western blot. Results The mRNA level of chemerin decreased significantly after four and ten weeks MCD feeding, although no obvious changes were found at first week, similar changes were found in serum chemerin (1.00±0.11 vs 0.96±0.39; 1.00±0.12 vs 0.21 ±0.77; 1.00±0.42 vs 0.21 ±0. 11). Contrasting with the change of chemerin(1.00±0.08 vs 0.72±0.10;1.00±0.24 vs 0.63±0. 31 ;1.00±0.05 vs 0.50±0.13), the mRNA level of CMKLR1 increased after MCD feeding( 1.00±0. 14 vs 0. 84±0. 26; 1.00±0. 38 vs 1. 51 ±0. 33; 1. 01 ±0. 13 vs 1. 84 ± 0. 39 ). Conclusion The change of chemerin and its receptor may participate in the process of the nonalcoholic fatty liver disease.

This study is to observe the protection of gross saponins of Tribulus terrestris (GSTT) on cardiocytes impaired by adriamycin (ADR) and approach its mechanism of action. Cardiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, model (ADR 2 mg x L(-1)) group, and GSTT (100, 30, and 10 mg x L(-1)) groups. MTT colorimetric method was deployed to detect cardiocyte survival rate, activities of CK, LDH, AST, SOD, MDA and NO were detected, and apoptosis was detected with flow cytometry. Effect of GSTT on caspase-3 was detected with Western blotting. Compared with control group, contents of CK, LDH, AST, MDA and NO were increased, and activity of SOD was reduced (P < 0.05, P < 0.01, P < 0.001) by ADR. Numbers of survival cells were increased (P < 0.05, P < 0.001), contents of CK, LDH, AST, MDA and NO were decreased, and activity of SOD was increased (P < 0.05, P < 0.01, P < 0.001) by GSTT (100 and 30 mg x L(-1)). Apoptosis of cardiocytes and concentration of caspase-3 can be reduced by GSTT (100 and 30 mg x L(-1)). GSTT can protect cardiocytes impaired by ADR, which are possible involved with its effect of resisting oxygen free radical.
Animals ; Antibiotics, Antineoplastic ; toxicity ; Apoptosis ; drug effects ; Aspartate Aminotransferases ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; drug effects ; Cells, Cultured ; Creatine Kinase ; metabolism ; Doxorubicin ; toxicity ; L-Lactate Dehydrogenase ; metabolism ; Malondialdehyde ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Nitric Oxide ; metabolism ; Rats ; Rats, Wistar ; Saponins ; isolation & purification ; pharmacology ; Superoxide Dismutase ; metabolism ; Tribulus ; chemistry