Chloroplast genetic engineering, offers several advantages over nuclear transformation, including high level of gene expression, increased biosafety, remedying some limitations associated with nuclear genetic transformation, such as gene silencing and the stability of transformed genes. It is now regarded as an attractive new transgenic technique and further development of biotechnology in agriculture. In this article we reviewed the characteristics, applications of chloroplast genetic engineering and its promising prospects were discussed.
Biotechnology ; methods ; Chloroplasts ; genetics ; Genetic Engineering ; methods ; Plants, Genetically Modified ; genetics ; Transformation, Genetic

OBJECTIVETo establish an animal model of tumor-associated depression and observe their biological behaviors and biochemical indices.

METHODSFour groups of SD rats kept in separate cages were subjected to tumor cell inoculation with or without chronic unpredictable moderate stress administered before or after the inoculation. The depressive behaviors of the rats were examined by open-field test, and the concentration of 5-HT in the hippocampus was measured by spectrophotofluorometry. The body weight of the rats and volume of the implanted tumor were monitored and sugar water test was performed.

RESULTSThe rats subjected to chronic stress displayed significant depression, manifested by reduction in movement in the central area and total movement distance with prolonged resting time and shortened time of activity. These rats maintained high levels of depression even 12 days after withdrawal of chronic stress. Compared with the control group, the depressive rats showed obviously reduced sugar water consumption and hippocampal 5-HT level. Tumors of different sizes were observed in all rats in the 4 groups.

CONCLUSIONA rat model of tumor-associated depression is established, and the tumor-bearing rats exhibit obvious depressive behaviors and reduced level of neural substance (5-HT), which provides a good basis for studying the association of depression with tumorigenesis,progression and prognosis of tumor.

Animals ; Carcinoma 256, Walker ; complications ; psychology ; Depressive Disorder ; etiology ; Disease Models, Animal ; Female ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stress, Psychological ; complications

BACKGROUNDThe treatment of high-grade developmental spondylolisthesis (HGDS) is still challenging and controversial. In this study, we investigated the efficacy of the posterior reduction and monosegmental fusion assisted by intraoperative three-dimensional (3D) navigation system in managing the HGDS.

METHODSThirteen consecutive HGDS patients were treated with posterior decompression, reduction and monosegmental fusion of L5/S1, assisted by intraoperative 3D navigation system. The clinical and radiographic outcomes were evaluated, with a minimum follow-up of 2 years. The differences between the pre- and post-operative measures were statistically analyzed using a two-tailed, paired t-test.

RESULTSAt most recent follow-up, 12 patients were pain-free. Only 1 patient had moderate pain. There were no permanent neurological complications or pseudarthrosis. The magnetic resonance imaging showed that there was no obvious disc degeneration in the adjacent segment. All radiographic parameters were improved. Mean slippage improved from 63.2% before surgery to 12.2% after surgery and 11.0% at latest follow-up. Lumbar lordosis changed from preoperative 34.9 ± 13.3° to postoperative 50.4 ± 9.9°, and 49.3 ± 7.8° at last follow-up. L5 incidence improved from 71.0 ± 11.3° to 54.0 ± 11.9° and did not change significantly at the last follow-up 53.1 ± 15.4°. While pelvic incidence remained unchanged, sacral slip significantly decreased from preoperative 32.7 ± 12.5° to postoperative 42.6 ± 9.8°and remained constant to the last follow-up 44.4 ± 6.9°. Pelvic tilt significantly decreased from 38.4 ± 12.5° to 30.9 ± 8.1° and remained unchanged at the last follow-up 28.1 ± 11.2°.

CONCLUSIONSPosterior reduction and monosegmental fusion of L5/S1 assisted by intraoperative 3D navigation are an effective technique for managing high-grade dysplastic spondylolisthesis. A complete reduction of local deformity and excellent correction of overall sagittal balance can be achieved.

Adolescent ; Adult ; Child ; Child, Preschool ; Decompression, Surgical ; methods ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Radiography ; Spinal Fusion ; methods ; Spondylolisthesis ; diagnostic imaging ; surgery ; Young Adult

BACKGROUNDDifferential diagnosis of isolated calf muscle vein thrombosis (ICMVT) and gastrocnemius hematoma is essential for early identification of deep vein thrombosis (DVT). This study aimed to investigate the diagnostic value of high-frequency color Doppler ultrasound for differential diagnosis of ICMVT and gastrocnemius hematoma.

METHODSA retrospective case series of 35 ICMVT (M:F, 21:14; mean age (64.5 ± 10.6) years) and 23 gastrocnemius hematoma (M:F, 16:7; mean age (75.4 ± 11.8) years) patients with bilateral/unilateral lower limb pain was conducted between January 2006 and September 2012. Characteristics and the morphology of high-frequency color Doppler ultrasonography of the lower limb deep vein, great saphenous vein, calf muscles, skin, and soft tissue were examined.

RESULTSICMVT hypoechoic signals were characterized by long, tube-like masses on longitudinal sections and oval masses on transverse sections, with apparent muscle thrombosis boundaries, distal and proximal venous connections, and, often, lower limb DVT. Gastrocnemius hematoma hypoechoic signals were characterized by large volumes, enhanced posterior hematoma echo, hyperechoic muscle boundaries, no hematoma blood flow, and no DVT, and clear differences in trauma/exercise- and oral anticoagulant-induced hematomas were readily apparent. According to the measurement, the ratio of long diameter/transverse diameter (D/T) in ICMVT patients was about less than 2.0, whereas in gastrocnemius hematoma patients the ratio was more than 2.0. Early stage isoechoic and hypoechoic signals were detected with gradually increasing ovular anechoic areas. Partial muscle fibers in the hematoma due to muscle fractures were apparent.

CONCLUSIONHigh-frequency color Doppler ultrasound was found to be a sensitive and reliable method for differential diagnosis of ICMVT and gastrocnemius hematoma due to trauma and exercise or prolonged oral anticoagulant use.

Aged ; Diagnosis, Differential ; Female ; Humans ; Leg ; diagnostic imaging ; Male ; Middle Aged ; Muscle, Skeletal ; diagnostic imaging ; Retrospective Studies ; Ultrasonography ; Venous Thrombosis ; diagnostic imaging

OBJECTIVETo study the effect of ex vivo expansion on the adhesion activities of umbilical cord blood hematopoietic stem and progenitor cells (HSPC).

METHODSFresh UCB CD(34)(+) cells were cultured in a serum and stroma-free culture system. At day 7, day 10 and day 14, CD(34)(+) cells were re-selected from the expanded products. The expression of adhesion molecules (CAMs) such as VLA-4, VLA-5, LFA-1, ICAM-1, HCAM, L-selectin and PECAM-1, and the adhesion activity of the expanded CD(34)(+) cells were evaluated and compared with those of precultured fresh CD(34)(+) cells.

RESULTS(1) The CD(34)(+) cells expressing homing-related CAMs were increased (from 15-fold increase for CD(34)(+) CD(54)(+) subset to 72-fold increase for CD(34)(+) CD(49e)(+) subset at day 14). (2) The expressions of CD(49d), CD(44), CD(11a) and CD(49e) on the expanded CD(34)(+) cells were increased or sustained the same levels as those on fresh UCB CD(34)(+) cells, while the expression of CD(62L), CD(54) and CD(31) on expanded CD(34)(+) cells declined with the cultivating. (3) Spontaneous adhesion and SDF-1-induced adhesion tended to be increased in the course of the first 10 day's culture.

CONCLUSIONSThe culture system used in this study could substantially support the expansion of HSPCs expressing the above CAMs, and the expanded HSPCs would sustain their intrinsic adhesion potentials.

Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Cell Adhesion ; Cell Adhesion Molecules ; biosynthesis ; Cell Division ; Fetal Blood ; cytology ; immunology ; metabolism ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; immunology ; metabolism ; Humans ; Receptors, Lymphocyte Homing ; biosynthesis

OBJECTIVETo evaluate the Angelica Sinensis as a protecting agent affecting the radiation-induced pulmonary fibrosis in an animal model,

METHODSThe thoraces of C57BL/6 mice were exposed to either sham irradiation or single fraction of 12 Gy. Four groups were defined: that received neither irradiation nor Angelica Sinensis (N group), that received Angelica Sinensis but no irradiation (A group), that underwent irradiation without Angelica Sinensis (NX group) and that received both Angelica Sinensis and irradiation (AX group). Mice were sacrificed at 1, 24, 72 hours and 1, 2, 4, 8, 16, 24 weeks post-irradiation. The lungs tissue were removed and processed for definitive analysis, including hydroxyproline content, HE and Masson staining, and the TGF-beta1, (Transforming Growth Factor beta1, TGF-beta1) mRNA expressions.

RESULTSCompared with N and A group, there was some differences in the AX group, but a significant histological and pathologic changes in NX group. Non-irradiated groups (N and A group) exhibited low levels of hydroxyproline (0.775 +/- 0.024) microg/mg and (0.751 +/- 0.034) microg/mg, and there was a significantly elevated level of hydroxyproline in NX group (0.875 +/- 0.009) microg/mg (P < 0.05). AX group (0.782 +/- 0.010) microg/mg was in between the non-irradiated groups (N and A group) and the radiation-only group (NX group), and the difference between AX group and NX group was significant (P < 0.01). The results of real-time quantitative RT-PCR showed that the relative mRNA expressions of cytokine TGF-beta1 in NX group(249.655 +/- 16.320) was significantly higher than that in group A (1.254 +/- 0.061) and N (1.324 +/- 0.057) (P < 0.01), and that in AX group (108.076 +/- 9.870) decreased than that of NX group (P < 0.01).

CONCLUSIONAn animal model of mice with radiation-induced lung injure was established successfully. This study has demonstrated that Angelica sinensis in Hibits the progress of radiation-induced pulmonary fibrosis, Possibly by down-regulating the expression of the proinflammatory cytokine Tgfb1. These data suggest that Angelica sinensis maybe useful in preventing and/or treating radiation-induced pulmonary fibrosis in the clinic.

Angelica sinensis ; Animals ; Female ; Mice ; Mice, Inbred C57BL ; Phytotherapy ; Pulmonary Fibrosis ; drug therapy ; etiology ; pathology ; Radiation Injuries, Experimental ; drug therapy ; pathology ; Radiation-Protective Agents ; Transforming Growth Factor beta1 ; metabolism

AIM To study the effects of Yizhi Congming Decoction (Ginseng Radix et Rhizoma,Poria,Polygoni multiflori Radix praeparata,etc.) on learning and memory deficits through reducing tau hyperphosphorylation in Aβ-induced AD mice and its mechanism of action.METHODS Sixty ICR mice were randomly and equally assigned into six groups.Sham group (injection of 3 μL normal saline into left hippocampus),model group,donepezil group and each group of Yizhi Congming Decoction (injection of 3 μL Aβ25-35 into left hippocampus).Donepezil group and three treatment groups were gavaged with donepezil and Yizhi Congming Decoction.After 15 days of administration,immunohistochemistry was used to observe the expressions of Ser404 and Thr231.Western blot was performed to evaluate the levels of phosphatidylinositol-3-kinase (PI3K),threonine/serine protein kinase B (AKT) and glycogen synthase kinase 3β (GSK3β).RESULTS Yizhi Congming Decoction attenuated the levels of phosphorylated tau at the Thr231 and Ser404 sites in the hippocampus and increased the phosphorylation levels of PI3K,AKT and GSK3β.CONCLUSION Yizhi Congming Decoction effectively provides protection against learning and memory deficits and inhibits hyperphosphorylated tau protein expression in the hippocampus.The possible mechanism may occur via the PI3 K/Akt-dependent GSK3 β signalling pathway.

Objective:To study the safety and efficacy of doxofylline for treating the patients with ticagrelor caused dyspnea.Methods:A total 172 coronary artery disease (CAD) patients with ticagrelor caused dyspnea in our hospital from 2015-02 to 2016-07 were studied.The patients were divided into 2 groups:Intervention group,patients received doxofylline at 200 mg twice per day for 5 days and Control group,patients received placebo.n=86 in each group.Dyspnea remission rate of was recorded at 1 day after treatment;platelet aggregation rate before and after treatment,cardiac death,myocardial infarction (MI),stroke,bleeding and other adverse cardiovascular and cerebral events were compared at 6 month after treatment.Results:Compared with Control group,Intervention group had improved dyspnea remission rate at 1 day after treatment (93％ vs 63％),P＜0.05;platelet aggregation rate [before doxofylline application:(35.53±5.1)％ vs (35.16±4.6)％,after doxofylline application:(26.48±4.3)％ vs(25.98±4.7)％]adverse cardiovascular and cerebral events were similar between 2 groups before and after doxofylline application,P＞0.05.Conclusion:Doxofylline was effective for treating the patients with ticagrelor caused dyspnea,it does not affect platelet aggregation effect of ticagrelor.

Objective:To explore the miRNA (micro RNA)differential expression profile between primary retroperitoneal liposarcoma tissues and normal fat tissues,and to provide the evidence that miRNA were involved in the molecular pathways of primary retroperitoneal liposarcoma tissues' occurrence.Methods:Collecting retroperitoneal liposarcoma tissues and normal fat tissues from 4 patients after radical surgery of retroperitoneal lipsarcoma.Using microarray analysis.The tissues' miRNA hybridizated with human's LC Sciences microRNA Microarray-Single (miRBase 21.0) expression profile gene chips,and got the date.Analyzing the differential expressing of the siginal date by LOWESS.Results:Total 38 differential expressed miRNA were found (P＜0.05),including 23 over-expression and 15 low-expression miRNAs.10 of them(38 differential miRNAs) was significant deviation (P＜0.01),including 4 over-expression and 6 low-expression.Date analysis revealed that some miRNAs were associated some different tumors,Conclusion:The number of over-expression were more than the low-expression in primary retroperitoneal liposarcoma compared with the normol fat tissue,which indicate that the genes expression are less abundant in primary retroperitoneal li-posarcoma;some of the miRNAs might involved in the molecular pa-thways of primary retroperitoneal liposarcoma tissues' occurrence and recurrence,they might become the target point of the targetedtherapy of the primary retroperitoneal liposarcoma,some of the over-expressed miRNAs can become new biomarkers in the following diagnosis of the primary retroperitoneal liposarcoma.

Objectives: To compare regional cerebral tissue oxygen saturation (rScO2) changes during cardiac surgery in children with ventricular septal defect (VSD) and tetralogy of Fallot (TOF). Methods: A total of 60 children aged 3~36 months were enrolled (28 in VSD group 32 in TOF group). rScO2was monitored by Fore-Sight near-infrared spectroscopy device. rScO2, pulse oxygen saturation (SpO2), systolic pressure, diastolic pressure, heart rate, stroke volume index, cardiac index, systemic vascular reststance index the maximal slope of systolic upstroke (dp/dt max) were obtained at following time points: after anesthesia induction (t0), pericardium opening (t1), 5 min after cardiopulmonary bypass (CPB) initiation (t2), 5 min before separation from CPB (t3), separation from CPB (t4), post-modified ultrafiltration (t5), end of surgery (t6). Results: (1) The lowest rScO2value was observed at separation from CPB (t4), and which was significantly lower than that at t0(P<0.05) for both groups; rScO2, stoke volume index, cardiac index, and dp/dt max at t5were significantly higher than at t4(all P<0.001) for both groups. (2) rScO2and SpO2were significantly higher at t5and t6than at t0(both P<0.05) in TOF group. rScO2at t0-t2was significantly lower in TOF group than in VSD group (both P<0.05). rScO2increased more significantly after modified ultrafiltration and rScO2was positively correlated with SpO2at t0and t1(r=0.35, P<0.05 and r=0.64,P<0.01) in TOF group. (3) In the total cohort, rScO2was positively correlated with age, weight at t0, t1, t3, t4, t5, t6(all P<0.01). After modified ultrafiltration, the increase in cardiac index was positively correlated with increase in rScO2(r=0.41, P<0.05), and the amount of cardiac index and rScO2increases were negatively correlated with age (r=-0.30;r=-0.34, both P<0.05). Conclusions: rScO2is closely related with age and weight. Cerebral oxygen delivery before biventricular surgical correction is lower in TOF group than in VSD group, and the cerebral oxygenation improves significantly after surgical repair. Modified ultrafiltration could significantly improve systemic hemodynamics and rScO2, and TOF children and younger children benefit more from modified ultrafiltration. Pre- and post-separation from CPB period is vulnerable to cerebral desaturation, it is therefore of importance to maintain the cerebral oxygen delivery-consumption balance at these periods for children with TOF and VSD undergoing surgical repair.