RT-PCR amplification of ginseng ?-amyrin synthase gene was successfully performed based on the total RNAs extracted from ginseng hairy roots induced by Agrobacterium rhizogenes A4 using a modified guanidine isothiocyanate-method. Sequence analysis of this gene revealed that its sequence was consistent with the sequence of a previously reported ginseng ?-amyrin synthase gene (GenBank No. AB009030). This gene was inserted into pMD-119T simple vector and transformed into E.coli DH5?. Furthermore antisense plant expression vector of this gene was constructed using the pBI121 vector, laying foundation for studies on antisense regulation of ginseng ?-amyrin synthase gene.

Genetic engineering is a powerful tool in Panax ginseng breeding.Genetic transformation and plant regeneration are the premise and foundation involved in genetic engineering of Panax ginseng.Ginseng can be regenerated through organogenesis or somatic embryogenesis and indirect somatic embryogenesis is mainly used for its regeneration.Summurized the factors influencing plant regeneration such as different explants,different carbohydrates,somatic embryo optimization and hormone-free approach.Ginseng transformation has been achieved by Agrobacterium tumefaciens and Agrobacterium rhizogenes and transgenic ginseng with good characters was obtained by introducing genes associated with biosynthesis of ginsenosides or herbicide gene.Hairy root culture system can supply large scale of ginsenosides,thus effect of rolC genes on ginseng hairy root induction,regeneration and bioreactor culture of hairy root were discussed.Additionally,problems that are present in genetic engineering of Panax ginseng were also discussed in this review.

Objective To explore the protective effect of neuregulin-1 (NRG-1) on heart function and myocardium in rats with sepsis and its mechanism . Methods Healthy male Sprague-Dawly (SD) rats were divided into three groups according to random number table method, with 6 rats in each group. Sepsis model was established by cecal ligation and puncture (CLP group); rats in sham operation group (sham group) underwent the same procedure except ligation. Rats in NRG-1 pre-treatment group (NRG group) were intravenously injected with recombinant human NGR-1 (rhNRG-1) at the dose of 10 μg/kg through tail vein; rats in CLP group and sham group were treated with the same amount of saline. At 24 hours after CLP, hemodynamic method was used to evaluate the cardiac function, and myocardial morphology was observed with hematoxylin and eosin (HE) staining, enzyme linked immunosorbent assay (ELISA) was used to detect the levels of cardiac troponin T (cTnT), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) in serum and macrophage migration inhibitor factor (MIF) in myocardial tissue. Results ① heart function: compared with the sham group, the mean arterial pressure (MAP), left ventricular systolic pressure (LVSP) and left ventricular pressure maximal rate of rise and fall (±dp/dt max) were significantly decreased in CLP group and NRG group, while the MAP, LVSP and ±dp/dt max in NRG group were significantly higher than those in CLP group [MAP (mmHg, 1 mmHg = 0.133 kPa): 125.78±8.15 vs. 113.05±5.85, LVSP (mmHg): 151.27±6.79 vs. 139.39±8.05, +dp/dt max (kPa/s): 4 389.59±332.38 vs. 3 706.85±451.31, -dp/dt max (kPa/s): 4 291.42±323.72 vs. 3 691.17±515.44, all 1 <0.05]. ②Myocardial injury: compared with the sham group, the levels of serum cTnT in CLP group and NRG group were significantly increased, while the levels of serum cTnT in NRG group were significantly lower than those in CLP group (ng/L: 206.37±67.28 vs. 344.13±80.95, 1 < 0.05), and the HE staining showed that myocardial pathological changes in NRG group were improved compared with the CLP group. ③Inflammatory mediators level: compared with the sham group, the levels of serum TNF-α, IL-1β and myocardial MIF were significantly increased in CLP group and NRG group, while the indicators in NRG group were lower than those in CLP group [TNF-α(ng/L): 52.77±3.43 vs. 97.19±13.98, IL-1β (ng/L): 40.25±5.48 vs. 56.05±6.88, MIF (μg/L): 1.92±0.16 vs. 2.87±0.10, all 1 <0.05]. Conclusion NRG-1 can reduce circulating levels of inflammatory factors in rats with sepsis, adjust myocardial MIF level, and alleviate myocardial cell injury, thereby improving cardiac function, and play a role in myocardial protection.

Objective To optimize the extraction method of Coptidis Rhizoma protein; To compare and analyze the differences of protein in different parts of wild and cultivated Coptidis Rhizoma. Methods SDS-PAGE gel electrophoresis and Coomassie brilliant blue method were used to compare the differences of the protein of Coptis chinensis from the 3 extraction methods (water extraction, Tris-HCl and ammonium sulfate precipitation), and the protein in different parts of the wild and cultivated Coptis chinensis, and analyzed and evaluated the differences. Results The optimum extraction method of Coptis chinensis protein is Tris-HCl method, and the protein content of Rhizoma Coptidis is the highest, and the content of protein is the highest in every gram of medicinal material. The differences among different parts of wild and cultivated Coptidis Rhizoma were obvious, and the ranks for protein abundance and protein content per gram were: rhizome of cultivated Coptis chinensis＞ rhizome of wild Coptis chinensis＞ stem and leaf of cultivated Coptis chinensis＞ stem and leaf of wild Rhizoma Coptidis＞ fibrous roots of cultivated Coptidis Rhizoma. Cluster analysis showed that the correlation between protein of wild and cultivated rhizome of Coptidis Rhizoma was obvious. Conclusion The optimum Tris-HCl method can extract the protein from Coptidis Rhizoma, and the protein content of roots of Coptidis Rhizoma is significantly higher than other parts.

Nowadays,with the continuous deepening and development of vocational education teaching reform,medical higher vocational education always takes moral education as the fundamental task.As an independent type of education,vocational education should always deepen the integration of industry and education and the integration of science and education.Through the teaching research of"problem chain+course ideological and political case",this study innovates the coordinated education team of drug nursing curriculum,the collaborative education method and the collaborative education evaluation,and improves the teaching effect.

OBJECTIVETo compare and analyze the clinical effects of external fixator and small splint fixator in the treatment of comminuted distal radius fracture in senile.

METHODSFrom 2005.6 to 2008.6, 74 senile patients (82 sides) with comminuted distal radius fractures were divided into external fixation group (34 cases 38 sides, 27 males and 7 females, with an average of 70.05 +/- 3.70 years) and small splint fixation group (40 cases 44 sides, 29 males and 11 females, with an average of 70.30 +/- 3.48 years). The loss of volar tilting angle and ulnar inclination angle after reduction and the function scores of carpal joint after removing the fixators were compared.

RESULTSOne week after surgery, there was loss of volar tilting angle and ulnar inclination in small splint fixation (P < 0.01), and one month after removing the external fixator, the loss of angle was more obvious (P < 0.01); while the loss of angle in external fixation group was not significant (P > 0.05). After one month of removing the fixation, the functional score of wrist joint in external fixation group was obviously higher than that of the small splint fixation group (P < 0.05).

CONCLUSIONThe external fixator can be adopted to treat comminuted distal radius fractures in senile, which is able to decrease the reduction loss and helpful to functional recovery.

Aged ; Case-Control Studies ; External Fixators ; Female ; Humans ; Male ; Radius Fractures ; surgery ; Treatment Outcome

OBJECTIVETo investigate the effectiveness of surgical approaches, outcomes and prognosis of aortic root pathology due to Stanford A aortic dissection.

METHODSRetrospective analysis the clinical data of 161 patients (122 male and 39 female, mean age of (44 ± 21) years) underwent surgical treatment for Stanford A aortic dissection between January 2001 and June 2011. There were 146 patients of acute aortic dissection and 15 patients of chronic aortic dissection. All the patients had aortic root pathologies that included commissural prolapsed in 140 cases, more than moderate aortic insufficiency in 75 cases, aortic sinus intima rupture in 15 cases, right and/or left coronary artery tearing in 8 cases, right and/or left coronary artery dissection in 16 cases, aortic root aneurysm in 31 cases.

RESULTSAortic root replacement (Bentall procedures) were used in 72 cases, aortic root remodeling (including aortic valve replacement) in 80 cases, aortic root reimplantation (David procedure) in 9 cases. The cardiopulmonary bypass time was shorter in aortic root remodeling group ((193 ± 42) minutes) than the other two groups ((210 ± 61) minutes, (197 ± 34) minutes, F = 3.22, P = 0.04). The in-hospital mortality was 8.1% (13 cases), 5 cases (6.9%) in aortic root replacement group, 7 cases (8.8%) in aortic root remodeling group, 1 case in aortic root reimplantation. The cause of death included respiratory failure (4 cases), permanent neurological deficits (3 cases), multiple organ failure (4 cases), acute renal failure (2 cases). The survivors were followed up for 6 months to 6 years. There was no patient required reoperation for aortic root pathologies. There was no statistically significant difference between aortic root remodeling group and reimplantation group (P > 0.05).

CONCLUSIONSThe surgical treatment for aortic root pathology due to Stanford A aortic dissection is challenging. Appropriate procedures according to the specialty of aortic root pathology can be performed with favorable functional results.

Adolescent ; Adult ; Aged ; Aneurysm, Dissecting ; surgery ; Aorta ; pathology ; surgery ; Aortic Aneurysm ; surgery ; Aortic Valve ; surgery ; Blood Vessel Prosthesis Implantation ; methods ; Female ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVE: To determine the influence and significance of intermittent ventilation on lung cytokines during cardiopulmonary bypass (CPB). METHODS: Thirty-six patients with rheumatic heart disease (RHD) were divided into 2 groups randomly: Treatment group (n = 18, given intermittent ventilation once every 5 minutes during CPB; control group ( n = 18), no ventilation during CPB. A brochioalveolar lavage was performed at 2 hours after the CPB. The numbers of granulocytes and the contents of IL-1, IL-6, TNF-alpha, and LDH in the brochioalveolar lavage fluids were measured, and lung PaO2/FiO2 were measured preoperatively and at 1 hour, 4 hours, and 8 hours after the CPB termination. RESULTS: The numbers of granulocytes and the contents of IL-1, IL-6, TNF-alpha, and LDH of the treatment group in the brochioalveolar lavage fluids were significantly lower than those of the control group (P < 0.05), and the lung PaO2/FiO2 of the treatment group at 1 hour, 4 hours, and 8 hours after the CPB termination significantly increased than those of the control group (P < 0.05). A significant decrease of lung PaO2/FiO2 occurred in both groups at 1 hour, 4 hours, and 8 hours after the CPB compared with the same group at the baseline before the CPB (P < 0.01). CONCLUSION Intermittent ventilation has protective effects on the lung injury during CPB by decreasing granulocyte adhesion and the level of lung cytokines, alleviating the lung inflammatory reaction and endothelial cell injury.
Adult ; Cardiopulmonary Bypass ; Cytokines ; metabolism ; Female ; Humans ; Interleukin-1 ; metabolism ; Interleukin-6 ; metabolism ; Intermittent Positive-Pressure Breathing ; Lung ; metabolism ; Male ; Middle Aged ; Rheumatic Heart Disease ; physiopathology ; surgery ; Tumor Necrosis Factor-alpha ; metabolism

In order to elucidate the effect of acetylcholine (ACh) on the occurrence and development of human pituitary adenoma, it was firstly observed whether there exists choline acetyl transferase (ChAT) which is necessary for the synthesis of acetylcholine in the cells of human pituitary adenoma, and then MTT method, (3)H TdR incorporation, cell cycle analysis and TUNEL were employed to estimate the influence of ACh on the proliferation, DNA synthesis and apoptosis of three kinds of human pituitary adenoma (human prolactinoma, somatotropinoma and non-functional tumor) cells cultured in vitro. The results showed that (1) the positive staining of ChAT was obviously observed in the cells of the three kinds of human pituitary adenoma, however, it was lower than that in normal human pituitary gland; (2) ACh had a similar effect on the proliferation of the three kinds of human pituitary adenoma cells. ACh at 0.1-10 micromol/L decreased the (3)H TdR incorporation and the MTT A value in a dose-dependent manner. At the same time, ACh decreased the ratio of S or G(2) phase pituitary adenoma cells significantly, but increased the ratio of G(1) phase pituitary tumour cells markedly; (3) the effect of acetylcholine on the proliferation of human pituitary adenoma cells was inhibited by atropine, but not by tubocurarine; (4) ACh had no effect on the apoptosis of human pituitary adenoma cells cultured in vitro. These data suggest that ACh may have a significant modulating effect on the proliferation of pituitary adenoma cells by means of paracrine or autocrine, and the effect is mediated by muscarinic receptor.
Acetylcholine ; pharmacology ; physiology ; Acetyltransferases ; biosynthesis ; physiology ; Adenoma ; pathology ; secretion ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Pituitary Neoplasms ; pathology ; secretion ; Tumor Cells, Cultured

To establish a systematic site-specific metastatsis model of human hepatocellular carcinoma (HCC) in nude mouse. HCCLM3-R cells were seeded into mice liver to establish xenograft mouse models. With the help of RFP, metastasis foci in lungs and lymph nodes in mice were detected using fluorescent stereomicroscopy and were removed. Cells derived from the metastasis foci were named HCCLM3-R-LM1 and HCCLM3-R-LnM1 respectively. HCCLM3-R-LM1 and HCCLM3-R-LnM1 cells were seeded into mice livers to analyze the lung and lymph node metastasis. Lungs of all tested mice were collected, examined by pathological evaluation and counted lung metastasis. Both lung and lymph node metastasis were found in HCCLM3-R-LM1, HCCLM3-R and HCCLM3-R-LnM1 cells and a significant difference was found between the lung and the lymph node metastasis levels in the three cells. The fluorescent areas (pixels) of lung and lymph node metastasis were 8687.00+/-1844.63 versus 2570.00+/-318.20 (P = 0.0031) in HCCLM3-R-LM1 cells, 6457.67+/-832.62 versus 10 994.33+/-2 212.31 (P = 0.0036) in HCCLM3-R cells, and 2968.67+/-2571.00 versus 24 416.00+/-7 186.13 (P = 0.0094) in HCCLM3-R-LnM1 cells, respectively. The middle numbers of microscopic lung metastatic foci were 775, 430 and 310 in HCCLM3-R-LM1, HCCLM3-R and HCCLM3-R-LnM1 cells (P less than 0.001), respectively, consist with the results quantified by RFP. We established the systematic site-specific metastasis models which demonstrates lung- and lymph node-specific metastasis potential in nude mice and can be used as a model for researches on site-specific metastasis of HCC.