To standardize the harvest ways of Dendrobium officinale and improve the quality and yield of D. officinale, a field experiment was carried out to study the effect of two kinds of harvest ways, which were keeping some of the axial shoot and harvesting all of the shoot by the end of the year. Then, the agronomic traits and yield were measured and the contents of polysaccharides and extractum were determined. The results showed that the harvest ways significantly affected the growth of D. officinale. Keeping some of the axial shoot could significantly improved the number of sprout, stem length, internode number and the internodal length, which also triggered increase the weight of fresh stems, leaves and the total of them and dry stems in per unit area, but it could not promote the stem diameter and the polysaccharide content in stems. Keeping some of the axial shoot moderately was conducive to the improvement of the production of medicinal materials in the process of harvesting by promoting the germination and growth of new buds, and to ensure the polysaccharide content by regulating the illumination and the density of cultivation.
Agriculture ; methods ; Dendrobium ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development

AIM: To study the causes of pyogenic granuloma after hydroxyapatite(HA) orbital implants.METHODS: HA orbital implants (250 cases) in our hospital (68 pegged implants) were reviewed.All patients were followed up from 18 months to 10 years. Implants were removed after medical therapy which was proved to be ineffective.RESULTS: Ten of 250 cases of HA orbital implants developed pyogenic granuloma. Pyogenic granuloma occurred in 1 unpegged implants patient and 9 patients after pegging and drilling of HA implantation over 4～7 years. The pyogenic granulomas were not controlled by medical therapy effectively. Implants were removed in 9 cases except 1 case denied removing and continued medical therapy.CONCLUSION: Pyogenic granuloma was serious complication that occurred after HA orbital implants. Partial vascularization, implant exposure, xenogenic sclera implant, pegging and drilling of HA implantation are risk factors that affect the development of pyogenic granuloma.Pyogenic granuloma hasn't relation with implanted peg material. Pyogenic granuloma denotes the potential implant infection, and all implants should be removed finally.

Objective:The recombinant human retinal pigment epithelium-derived factor(PEDF)protein to be obtained and the angiogenesis of the rPEDF to be identified.Methods: PEDF gene gene was amplified by PCR and cloned into pET32a,rPEDF protein was expressed in E.coli BL21 and confirmed by SDS-PAGE and Western blot.The rPEDF was purified by Ni-NTA on denature condition.The concentration of the rPEDF was determined by Bradford method.The angiogenesis of the rPEDF was determined by chick chorioallantoic membrane(CAM) method.Results: The expression plasmid pET32a-PEDF was constructed successfully.The rPEDF was expressed with stable efficiency in E.coli BL21.The results of the CAM experiment showed that the rPEDF had notable angiogenesis effect in the concentration 0.4、0.04 ng/ml,but had no effect in 4 ng/ml.Conclusion:The PEDF gene was cloned and expressed efficiently,the angiogenesis of the rPEDF to be identified and the activity was worked in certain range.The results can facilitate studying its function and spreading its application.

The study is aimed to make the most of available space of Dendrobium officinale cultivation facility, reveal the yield and functional components variation of stereoscopic cultivated D. officinale, and improve quality, yield and efficiency. The agronomic traits and yield variation of stereoscopic cultivated D. officinale were studied by operating field experiment. The content of polysaccharide and extractum were determined by using phenol-sulfuric acid method and 2010 edition of "Chinese Pharmacopoeia" Appendix X A. The results showed that the land utilization of stereoscopic cultivated D. officinale increased 2.74 times, the stems, leaves and their total fresh or dry weight in unit area of stereoscopic cultivated D. officinale were all heavier than those of the ground cultivated ones. There was no significant difference in polysaccharide content between stereoscopic cultivation and ground cultivation. But the extractum content and total content of polysaccharide and extractum were significantly higher than those of the ground cultivated ones. In additional, the polysaccharide content and total content of polysaccharide and extractum from the top two levels of stereoscopic culture matrix were significantly higher than that of the ones from the other levels and ground cultivation. Steroscopic cultivation can effectively improves the utilization of space and yield, while the total content of polysaccharides and extractum were significantly higher than that of the ground cultivated ones. The significant difference in Dendrobium polysaccharides among the plants from different height of stereo- scopic culture matrix may be associated with light factor.
Cell Culture Techniques ; methods ; Dendrobium ; chemistry ; growth & development ; Plant Leaves ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development ; Polysaccharides ; analysis

Adolescent ; Adult ; Aged ; Capsules ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis ; blood ; drug therapy ; Humans ; Male ; Middle Aged ; Phytotherapy ; Transforming Growth Factor beta ; blood ; Transforming Growth Factor beta1

Background Inherited retinal degeneration,one of the major causes of blindness worldwide,comprises a large number of disorders characterized by a slow and progressive retinal degeneration.Interleukin-1 (IL-1)was recognized to be involved in inherited retinal degeneration.Whether IL-1 receptor antagonist (IL-1ra) can arrest retinal degeneration is worthy of investigation.Objective This study was to investigate the effects of IL-1ra on photoreceptor apoptosis in Royal College of Surgeons (RCS) rats.Methods The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.The SPF RCS rats aged 9,15,16,25,30,35,40,50 and 60 postnatal days were collected,with 9 rats for each age group.Retinal sections were used for the TdT-mediated biotin-dUTP nick-end labeling (TUNEL) cell apoptosis assay.1 μl of IL-1ra (1.8 g/L) was intravitreally injected in the right eyes of 9 RCS rats aged 15 postnatal days and PBS was used in the same way in the fellow eyes.The injection procedure was repeated on the 20 th and 25 th day,respectively.The rats were sacrificed on the 30 th day and retinal sections were prepared for the TUNEL assay.The differences in the percentage of the positive cellular nuclei area among different ages of rats were compared by one-way ANOVA,and the differences in retinal layer thickness between IL-1ra injection group and PBS injection group were assessed by paired t test.Results Photoreceptor apoptosis appeared in 20-day-old RCS rats and progressed and peaked in 30 and 35-day-old rats and then reduced,showing a significant difference among rat of various age groups (F=28.020,P＜0.01).Images from TUNEL assay showed a weaker and less TUNEL staining in the IL-1ra injected eyes than the PBS injected eyes in 30-day-old rats.The total area and relative area of TUNEL positive nuclei were (223.052±153.092) μm2 and (2.206±1.531) ％ in the IL-1ra injected group,and those in PBS injected group were (1235.050±359.767) μm2 and (7.269± 1.624) ％,with a significant difference between them (t =4.370,t=3.250,P＜0.01).The cone and rod thickness was (15.324±9.035) μm in the IL-1ra injected group and (49.566±4.605)μm in the PBS injected group,showing a significant difference (t =22.674,P＜0.01).However,no significant difference was seen in the outer nuclear layer thickness between the two groups (t =0.268,P＞0.05).Conclusions IL-1 participates in the pathogenesis and development of inherited retinal degeneration of RCS rats.The use of IL-1ra might be a potential approach in the treatment of inherited retinal degeneration.

· AIM: To study the clinical characteristics of metastatic bacterial endophthalmitis.endophthalmitis were retrospectively studied.manifestation was found in 17 cases (77%), iridal purulent nodules in 2 cases and retinal purulent embolism In 3 cases. The total number of leucocytes and neutrophilic granulocytes was elevated markedly, while lymphocyte count was reduced significantly in early stage. Positive rate of bacterial cultivation was 36%.dophthalmitis often have apparent extraocular infected situation before eye Is involved. Ocular appearance and laboratory examination have some specificity in early stage, while bacterial cultivation, as a golden standard,needs to be further improved.

Nowadays,small peptides are always expressed in the form of fusion protein.The expression product contains many superfluous amino acids which can affect the biological functions of small peptides even expressed by GST fusion protein expression system.SUMO protease can cut SUMO fusion protein expressed by fusion expression system without any amino acid residues left on target protein thus become a hot topic in this field.Recombinant His-UlP1/pET3c/BL21(DE3)engineering strain was constructed by genetic engineering technology and the expression conditions were optimized in shake flaks.The process of high density fermentation was explored and different purification conditions were detected by chromatography.The results showed that SUMO protease could be expressed well after inducing the engineering strain by IPTG of 1.0mmol/L at 30℃ for 6 hours.The expression level of the strain in fermentation pots could reach 24.3% analyzed by SDS-PAGE.The purity of SUMO protease was more than 98% after further purification by cation exchange chromatography.The yield was 355mg SUMO protease per liter fermentation liquid.Western blot analysis demonstrated that there were immune reactions between IlP1 and 6?His antibodies,so it has established a good foundation for large-scale industralazation in the future.

OBJECTIVETo analyze the variation of serum insulin levels in critically ill children and investigate the underlying mechanism and clinical significance to provide the basis for treatment.

METHODTotally 332 critically ill children admitted in pediatric intensive care unit (PICU) of Hunan Children's Hospital from Nov., 2011 to April, 2012 were studied. The high insulin group (n = 332) was defined as insulin levels within 24 h > 11.1 mU/L and was divided into 2 groups: mildly elevated group (n = 194): 11.10 - 33.30 mU/L, increased three times group (n = 138): > 33.3 mU/L. Insulin, C-peptide and blood glucose were measured within 24 hours after admission, on day 3 and 7. Other results of inflammatory markers, lactate, cardiac enzymes, amylase, pancreatic ultrasound, hepatic and renal function as well as indicators related to severity and prognosis were recorded after admission.

RESULTThe peak of insulin level was seen on day 1, then presented a downward trend and reached the normal level on day 7. The peaks of blood glucose and C-peptide level were seen on day 1 then declined, the levels on day 7 were still slightly higher than normal level. The insulin level on admission (41.47 ± 30.85) mU/L were positively correlated with lactic acid (2.29 ± 1.81) mmol/L and procalcitonin level (5.08 ± 6.70) ng/ml (r = 0.370, P = 0.000; r = 0.168, P = 0.002) (P < 0.01). The insulin level on admission in children with 1 organ failure (41.24 ± 22.60) mU/L or 2 or multiple organ failure (48.98 ± 22.17) mU/L was higher than that in children with non-organ failure (34.11 ± 29.84) mU/L (U = 1621.001, P = 0.000;U = 1300.000, P = 0.000) (P < 0.01). The insulin level on admission in death group (52.99 ± 32.34) mU/L was higher than that in survival group (32.85 ± 24.10) mU/L (U = 1585.000, P = 0.000) (P < 0.01). Ten cases in death group were complicated with pancreatic damage and the average insulin level on admission was (65.29 ± 50.53) mU/L.

CONCLUSIONThe high insulin level was correlated with the degree of inflammatory response, ischemia and hypoxia. The high insulin level in critically ill children was relevant to the pancreatic damage, the severity of the disease, organ dysfunction, and evaluation of prognosis.

Adolescent ; Blood Glucose ; metabolism ; C-Peptide ; blood ; Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Child ; Child, Preschool ; Critical Illness ; Female ; Humans ; Infant ; Infant, Newborn ; Insulin ; blood ; Intensive Care Units, Pediatric ; Male ; Multiple Organ Failure ; blood ; mortality ; Pancreas ; metabolism ; pathology ; Prognosis ; Protein Precursors ; blood ; Survival

BACKGROUND:The traditional two-dimensional culture system has been widely used in the in vitro culture of human tissue stem cells,but it cannot really simulate the three-dimensional physiological microenvironment in the body, which is not conducive to the study of the biological behavior of human stem cells. OBJECTIVE: To detect the effect of the bioactivity of Col-Tgel in human hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs)in vitro and in vivo,by constructing a three-dimensional culture system stimulating the physiological microenvironment of the body. METHODS:(1)In vitro co-culture:Green fluorescent protein labeled MSCs(MSCs-GFP)and human umbilical cord blood CD34+cells were co-cultured in Col-Tgel for 3 days (three-dimensional culture group). Human umbilical cord blood CD34+cells were cultured in Col-Tgel for 3 days as single culture group. MSCs-GFP and human umbilical cord blood CD34+cells were co-cultured in Transwell chamber for 3 days as two-dimensional culture group. Human umbilical cord blood CD34+cells were cultured routinely as control group. The percentage of CD34+CD38-CD45RA-CD90+cells in each group was measured by flow cytometry. In situ immunofluorescence staining was used to detect the activity of cells that were co-cultured in Col-Tgel.(2)In vivo transplantation:NOD/SCID mice subjected to 24-hour X-ray irradiation were divided into two groups: in experimental group, MSC-GFP cells were resuspended in Col-Tgel and transplanted into the tibia of NOD/SCID mice; in control group, MSCs-GFP were resuspended in PBS and transplanted into the tibia of NOD/SCID mice. The MSC-GFP growth in the bone marrow was detected by two-photon/confocal microscopy at 3 days post transplantation. RESULTS AND CONCLUSION: (1) After co-culture in Col-Tgel for 3 days, the percentage of CD34+CD38-CD45RA-CD90+cells in the three-dimensional culture group was 2.8 times that of the two-dimensional culture group, indicating that the MSCs significantly promoted the expansion of CD34+CD38-CD45RA-CD90+cells in the Col-Tgel. The percentage of CD34+CD38-CD45RA-CD90+cells in the three-dimensional culture group was increased by 4.5 times compared with the single culture group and increased by 1.5 times compared with the control group. Immunofluorescence staining showed that the cell viability of human MSCs and human umbilical cord blood CD34+cells was not affected after co-cultured in Col-Tgel for 3 days.In the in vivo transplantation experiment,MSC-GFP cells could survive in the medullary cavity.In summary, Col-Tgel provides a new strategy for stem cell culture and in vivo growth by forming a three-dimensional system similar to the physiological environment in vivo.