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

Objective To investigate the effect of leptin on the tolerance of cultured rat brain astrocytes to ischemia and hypoxia.Methods The brain astrocytes isolated from neonatal SD rats,after purification and identification,were incubated in serum-and glucose-flee medium in the presence of 5%CO2+95%N2 for 90 min to induce isehemic and hypoxic injury. RT-PCR was performed to detect the expressions of the leptin receptors Ob-Ra and Ob-Rb in the cells, and colorimetry was used to measure the content of malonaldehyde(MDA) and lactate dehydrogenase(LDH) activity in the cell supematant.The expression level ofglial fibrillary acidicprotein(GFAP)in the cells was detected with fluorescence immunocytochemistry.Results Ischemic and hypoxic exposure of the cells induced obvious cell necrosis.Compared with the cells without the exposure,significantly decreased Ob-Rb expression(0.52±0.01 vs 1.32±0.01,P<0.05)and increased MDA,LDH and GFAP levels(709.68±47.16 vs 516.13±29.08,3.94±0.36 vs 1.81±0.21,and 0.122±0.016 vs 0.057±0.006,respectively,P<0.05) occurred after the exposure,whereas the expression level of Ob-Ra underwent no significant changes(3.87±0.13 vs 3.96±0.24,P>0.05). Compared with the exposed cells,the leptin-treated cells showed a significant reduction in MDA levels(3.94±0.36 vs 3.19±0.25,P<0.05) with significantly increased GFAP expression(0.057±0.006 vs 0.109±0.008, P<0.05)after the exposure, and the cells maintained basically intact cell morphology.Conclusion With neuroprotective effects against ischemic neuronal injuries,leptin canimprove the tolerance of rat brain astrocytes to ischemia and hypoxia.

OBJECTIVETo summarize the mid-term effectiveness of Coflex non-fusion internal fixation treatment of degenerative lumbar disease.

METHODSFrom October 2008 to December 2010, a retrospective analysis was carried out on 39 patients (29 males and 10 females) diagnosed as degenerative lumbar disease and treated with Coflex interspinous dynamic device, who had been followed up for 1 year at least, the average age was 45.5 years (range, 23 - 67 years). The results were assessed by Japanese Orthopedic Association (JOA) scores, visual analogue scale (VAS) scores, Oswestry disability index (ODI) scores and SF-36 scores; and the range of mobility (ROM), intervertebral disc height of the responsible and adjacent segments were measured on X-film before the operation and at last follow-up. Observed the therapeutic effect of the patients and compared the effect on the patients of different body mass index (BMI) and different age by the One-way analysis of variance and paired t test.

RESULTSThe 39 patients were followed up for 30.9 months (range, 12 - 37 months). At the last follow-up, JOA, ODI, VAS and SF-36 scores were improved by 70% ± 12%, 54% ± 12%, 77% ± 10% and 51% ± 9%, and were statistically significant (t = -33.289, 26.448, 26.596 and -20.772, P = 0.00). Patients with BMI ≥ 25 kg/m(2) had lower improvement rates in the scores than those with BMI < 25 kg/m(2) (F = 10.561, 5.850, 5.651 and 6.519, P < 0.05). The patients were 50 years older or younger couldn't affected the improvement rates in the scores statistically (P > 0.05). There were no significant difference in remaining disc height (P > 0.05), except that the intervertebral disc height of L4-5 increased slightly compared with the preoperative (t = -2.819, P = 0.008). In addition to the ROM of L3-4, L5-S1 and L1-S1 were not significantly different from the preoperative(P > 0.05), the ROM of L4-5 were decreased (t = 12.598, P = 0.000).

CONCLUSIONSThe mid-term effectiveness of Coflex non-fusion interspinous fixation in treatment of degenerative lumbar disease is worthy of recognition, and Coflex combined with Isobar has advantages in the treatment of multi-segment degenerative lumbar disease.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Internal Fixators ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Stenosis ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the effect of intestinal ischemia/reperfusion (I/R) injury on leptin and orexin-A levels in peripheral blood and central secretory tissues, and investigate the roles of leptin and orexin-A in acute inflammatory responses.

METHODSAn intestinal I/R injury rat model was established, and the rats were grouped according to duration of the reperfusion time following a 60-min ischemia. Radioimmunoassay was used to examine the protein levels of leptin in the serum and adipose tissue, and the protein levels of orexin-A in the plasma and hypothalamus. Reverse transcriptase-polymerase chain reaction was also performed to detect the mRNA expressions of adipose leptin and hypothalamus orexin-A.

RESULTSCompared with that before injury, serum leptin level of 60-min ischemia with 30-min reperfusion (I60'R30') group decreased significantly and that of I60'R360' increased significantly. Compared with the sham-operation group (sham) after injury, serum leptin level of I60'R360' group increased significantly, and adipose leptin protein levels of I60'R30' and I60'R90' groups decreased significantly, whereas that of I60'R360' group increased obviously. Compared with sham group after injury, adipose leptin mRNA expressions of I60'R30', I60'R240' and I60'R360' groups all increased significantly, while that of I60'R150' showed significant decrease. No significant changes were noted in the protein levels of orexin-A either in the plasma or hypothalamus after I/R injury. In comparison with sham group after injury, hypothalamus orexin-A mRNA expressions of I60'R30' and I60'R90' groups showed gradual but significant decrease, and till 150 min of reperfusion, the expression reached its lowest, followed then by slow recovery at 240 and 360 min, though still remaining significantly lower than that of sham group.

CONCLUSIONLeptin and orexin-A have a time-dependent response to intestinal I/R injury, but the former appears to exhibit a faster response, and they may play a certain role in the metabolic disorders of acute inflammation.

Animals ; Female ; Inflammation ; blood ; genetics ; physiopathology ; Intestine, Small ; blood supply ; metabolism ; Intracellular Signaling Peptides and Proteins ; blood ; genetics ; Leptin ; blood ; genetics ; Male ; Neuropeptides ; blood ; genetics ; Orexins ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; blood ; genetics ; physiopathology ; Reverse Transcriptase Polymerase Chain Reaction

AIMTo explore the effect of rat myocardial ischemia/reperfusion injury on leptin levels in serum and myocardium, and discuss the role of leptin in myocardial ischemia/reperfusion injury.

METHODSA myocardial ischemia/reperfusion injury model of rats was established, serum lactate dehydrogenase (LDH) and leptin levels were detected, and histopathological changes and leptin expressions in myocardium were investigated by hematoxylin-eosin staining and immunohistochemistry, respectively.

RESULTSSerum LDH of ischemia and reperfusion groups increased significantly (P < 0.05), suggesting the model was successfully established and a certain degree of local myocardial injury was induced. Serum leptin of ischemia group (6.34 +/- 2.49) ng/ml was significantly lower than control group (7.50 +/- 2.93 ng/ml, P <0.05). Leptin levels recovered gradually after reperfusion, reached (8.32 +/- 1.74)ng/ml at 2 h after reperfusion, which recovered to the level before injury (8.38 +/- 2.56) ng/ml, and showed a trend to increase as reperfusion time was elongated. Immunohistochemistry results showed that as compared with sham-operation group, myocardial leptin protein expressions of the other four groups were all significantly lower (P < 0.01), and decreased in order by 45 min ischemia/1 h reperfusion, 45 min ischemia/3 h reperfusion, 45 min ischemia and 45 min ischemia/2 h reperfusion.

CONCLUSIONLeptin level in the blood decreases significantly at the early 45 min after myocardial ischemia/reperfusion injury, and its expression in myocardium also decreases significantly. There may be a certain relationship between the pathological injury of myocardium and the changes of leptin.

Animals ; L-Lactate Dehydrogenase ; metabolism ; Leptin ; blood ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; physiopathology ; Myocardium ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To explore the effect of family support service intervention on improving the rehabilitation of patients with severe mental disorders in community and the mental health status and family burden of family members. Methods Using multi-stage random sampling method, 100 patients who met the diagnostic criteria of severe mental disorders were randomly selected from two communities, and then 100 patients who met the diagnostic criteria of severe mental disorders were randomly matched according to gender, age and diagnosis in other communities into the control group. The control group and intervention group were set up strictly according to the inclusion criteria of patients and their families. Results The average age of the 200 groups was (48.27±12.67) years, and the average age of the family members was (63.61±13.19) years. After intervention, the activity dailyliving scale (ADL) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). After intervention, the social disability screening schedule (SDSS) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). After intervention, there was no significant difference in the morningside rehabilitation status scale (MRSS) score between the intervention group and the control group at all time points (all P>0.05). After intervention, the SCL-90(self-reporting inventory) scores of the mental health of the family members in the control group were higher than those in the intervention group at all times (all P<0.05). After intervention, the family burden scale of diseases (FBS) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). Conclusions The intervention measures did improve the rehabilitation effect of severe mental disorder patients in community and the psychological and family burden of family members. A professional family support service team should be established.

Adult ; Blood Coagulation ; Emergency Treatment ; Female ; Humans ; Intraoperative Period ; Liver Transplantation ; Male ; Middle Aged

Objective To evaluate the bioequivalent between the two kinds of glipizide tablets.Methods With two-cycle crossover trial was design, 24 healthy male subjects were given a single oral dose 5 mg of glipizide test preparation and reference preparation.Serum samples were collected at different time points before administration and after adminis-tration.The plasma concentrations were measured by HPLC -MS/MS method.The pharmacokinetic parameters and relative bioavailability were calculated by WinNonlin 6.1 software.Results The main pharmacoki-netic parameters of the test and reference glipizide tablets were as fol-lows: tmax were ( 3.40 ±1.54 ) and ( 3.71 ±1.30 ) h, Cmax were (471.88 ±108.10 ) and ( 480.58 ±132.63 ) μg · L-1 , t1/2 were (5.15 ±1.31) and(5.32 ±1.27)h, AUC0-t were (2805.71 ±592.61 ) and (2873.40 ±697.57 )μg· h-1 · L, respectively.F0-t and F0-∞ of test preparation were ( 105.77 ±27.84 )% and ( 105.28 ±27.63 )% re-spectively.Conclusion The glipizide tablets test and reference are bio-equivalent.

[Objective]To investigate the expression changes of miR-133b in methamphetamine(MA)-induced neuro-nal injury in PC12 cells and its regulative effects on cellular apoptosis.[Methods]PC12 cells were cultured and divided into control group and MA treated group.In MA treated group,PC12 cells were insulted with 800μmol/L MA in culture medium. The cellular injury of PC12 cells was observed under microscope. The cellular apoptosis was detected by Hoechst33342/PI double staining,and the expression level of miR-133b was examined by real-time quantitative PCR(RT-PCR). Further-more,miR-133b mimic and inhibitors were transfected into PC12 cells to analyze miR-133b's function in MA-induced cell apoptosis.[Results]The data showed that 800 μmol/L MA could induce obvious cellular injury,cause neurite shortened and increase the cell apoptosis. The RT-PCR data showed that the expression of miR-133b of PC12 cells treated with MA de-creased significantly.The apoptosis rate of PC12 cells decreased after transfection of miR-133b mimic,while increased after transfection of miR-133b inhibitors.[Conclusions]High concentration of MA causes neuron damage and induces neuronal apoptosis,and also decreases the levels of miR-133b expression. Whereas,overexpression of miR-133b can reduce the apoptosis of cultured PC12 cells.Thus,miR-133b plays a crucial role in MA mediated neurotoxicity.This study provides a theoretical basis for elucidating the mechanism of MA-induced neurotoxicity and may provide a new strategy for treating MA addiction.

OBJECTIVETo observe the expressions of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines and explore their relationship with the metastasis of human prostate cancer.

METHODSThe expressions and distribution of E-cadherin and beta-catenin in LNCaP and ARCaP cell lines (IF11 and IA8) were detected by Western blot and immunofluorescent staining.

RESULTSThe expression of E-cadherin was high in LNCaP, but absent in IF11 and IA8, while beta-catenin was expressed highly in IF11 and LA8, but lowly in LNCaP. Immunofluorescent staining showed that E-cadherin was mainly in the membrane of LNCaP, while beta-catenin both in the membrane of LNCaP and in the nuclei of IF11 and IA8.

CONCLUSIONE-cadherin and beta-catenin are differently expressed and distributed in prostate cancer cell lines with different characteristics of epithelial-mesenchymal transition (EMT), and the abnormal activation of the beta-catenin signal pathway may be involved in the EMT of prostate cancer cells.

Cadherins ; metabolism ; Cell Line, Tumor ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; beta Catenin ; metabolism