Objective To study the genetic diversity of Sinopodophyllum hexandrum so as to provide basic molecular genetic evidence for protecting and exploiting the resource of S.hexandrum.Methods Six populations of S.hexandrum were analyzed by DALP molecular markers.To make up the genetic diversity correlation data by PopGene 1.31 software and cluster by UPGMA method,and establish the dendrogram.The tree was subsequently visualized with Treeview software.Results Five primer groups were screened and a total of 150 DNA fragments were amplified,among which 104 were polymorphic,i.e.the percentage of polymorphic bands(PPB) was 69.33%,the average number of DNA polymorphic band amplified by each primer group was 20.8 and the average PPB was 14.22%.In the six populations,the total observed number of alleles(Na) was 1.693 3 and the average Na was 1.142 2;The total effective number of alleles(Ne) was 1.417 1 and the average Ne was 1.083 8;The total Nei′s gene diversity(H) was 0.244 3 and the average H was 0.048 8;The total Shannon′s information index(I) was 0.364 3 and the average I was 0.073 0;The total gene diversity(Ht) was 0.244 3 while the gene diversity within a population(Hs) was 0.048 7.The coefficient of gene differentiation(Gst) was 0.800 5 between populations,namely 80.05% genetic variation occurring between populations and the rest 19.95% within population,and estimated gene flow(Nm) was 0.124 6.Conclusion The result indicats that S.hexandrum has much larger genetic differentiation among populations and gene flow has been blocked.This might be a result of species breeding system and population habitat.

BACKGROUND: The application of adipose derived stromal cells to tissue engineering has been more and more popular around the world. Compatibility of scaffold material is the key point for its further research.OBJECTIVE: To determine the growth rules of rabbit adipose-derived stromal cells with polylactic-co-glycolic acid (PLGA).DESIGN, TIME AND SETTING: An in vitro study was performed at the Institute of Ophthalmology, Otolaryngology Hospital,Fudan University and Shanghai Tissue Engineering Center from September 2007 to March 2009.MATERIALS: Six female New Zealand rabbits aged six months were used for extraction of adipose-derived stromal cells. PLGA was provided by Sigma, USA.METHODS: Adipose tissue was harvested from the nape fat pad of the rabbits following anesthesia. Primary cultured cells were established using type I collagenase and cell cultures were maintained with DMEM containing 10% volume fraction of fetal bovine serum. Cells were passaged when 80% was confluent. The fourth passages of cells were utilized for the study. PLGA consisted of polylactic acid and polyglycolic acid as the ratio of 7:3, and the relative molecular mass was 104900.MAIN OUTCOME MEASURES: Initially, the adherent rate of cells to scaffold was detected. After one week co-culture, the scaffold bearing adipose-derived stromal cells labeled with Dio agent were investigated by fluorescence inverse microscope,scanning electron microscopy and laser scanning microscope.RESULTS: The best adherent rate of adipose-derived stromal cells with PLGA reached 99%. After one-week-incubation in vitro,cells of fiber-shaped or ovule-shaped proliferated well and exhibited stratified growth on the surface of PLGA scaffold. In addition,it also secreted visible extracellular matrices, which could be examined by scanning electron microscopic examination.Meanwhile, the adipose-derived stromal cells grew well and distributed equably inside the PLGA in terms of the investigation with laser scanning microscope.CONCLUSION: The compatibility of adipose-derived stromal cells to PLGA in vitro was well.

Objective To study the function and diurnal variation of hypothalamic-pituitary-adrenal (HPA) axis and hypothalamic-pituitary-thyroid(HPT) axis,and the relationship with clinical characteristics of di-urnal rhythm in depressed patients. Methods Forty-nine depressed patients met diagnostic DSM-Ⅳ criteria and thirty-eight normal controls were collected. The plasma levels of cortisol (CORT), adrenocorticotropic hormone (ACTH), T3, T4, thyroid-stimulating hormone (TSH) at 7 am and 7 pm were measured by radioimmunoassay, then the results concemed were compared among the depressed groups with and without diurnal variation and control group by multivariate analysis of variance. Results The plasma level of morning CORT, morning and evening TSH ((365.94±120.78) nmol/L; (6.24±2.47)μIU/ml; (6.68±2.42)μIU/ml; respectively) were respective-ly higher than those of control group((284.91±83.39) nmol/L; (3.82±1.75)μIU/ml;(4.01±1.69)μIU/ ml, P<0.05). The plasma levels of morning and evening T4 were significantly lower than those of control group(P <0.05). The CORT morning-evening difference variable distinguished between depressed and control group (P< 0.001),but no significant differences could be found between the depressed groups with and without diurnal varia-tion. There were no significant differences in the CORT, T3, T4, TSH morning-evening difference variable between various clinical subtypes of depressed patient (unipolar/bipolar; psychotic/nonpsychotic). Conclusion Abnormal CORT morning-evening difference variable was observed in the depressed patients compared with normal controls,and may be used as a special trait mark of depressed group.

Through the comparative analysis of undergraduate specialty settings of medical colleges and universities in minority areas in the different periods,this paper reveals the characteristics of undergraduate specialty settings.It is necessary to follow the demand for social and economic development,and adhere to the medical and national characteristics,promoting the durable development of undergraduate specialty settings of medical colleges and universities in minority areas.

OBJECTIVETo culture rat prostate glandular epithelial cells and study their barrier functions in vitro.

METHODSRat prostate glandular epithelial cells were cultured in vitro. The expression of the tight junction protein claudin-1 was determined by immunohistochemistry, the structure and composition of the epithelial cells observed under the inverted microscope and transmission electron microscope. The transepithelial electrical resistances (TEERs) were monitored with the Millicell system. The permeability of the prostate glandular epithelial cells was assessed by the phenol red leakage test.

RESULTSCompact monolayer cell structures were formed in the prostate glandular epithelial cells cultured in vitro. Immunohistochemistry showed the expression of the tight junction protein claudin-1 and transmission electron microscopy confirmed the formation of tight junctions between the adjacent glandular epithelial cells. The TEERs in the cultured prostate glandular epithelial cells reached the peak of about (201.3 ± 3.5) Ω/cm2 on the 8th day. The phenol red leakage test manifested a decreased permeability of the cell layers with the increase of TEERs.

CONCLUSIONThe structure and function of rat prostate glandular epithelial cells are similar to those of brain capillary endothelial cells, retinal capillary endothelial cells, and intestinal epithelial cells. In vitro cultured prostate glandular epithelial cells have the barrier function and can be used as a model for the study of blood prostate barrier in vitro.

Animals ; Cell Membrane Permeability ; Cells, Cultured ; Claudin-1 ; metabolism ; Electric Impedance ; Epithelial Cells ; pathology ; physiology ; ultrastructure ; In Vitro Techniques ; Male ; Microscopy, Electron, Transmission ; Phenolsulfonphthalein ; pharmacokinetics ; Prostate ; metabolism ; pathology ; Rats ; Tight Junctions

Objective To retrospectively study the risk factors of aortic arch calcificationand its influence on the survival prognosis of maintenance peritoneal dialysis patients. Methods One hundred seventy-seven cases of maintenance peritoneal dialysis patients were enrolled, including 66 cases of aortic arch calcification cases. Their general dialysis data were collected for the evaluation of dialysis adequacy and residual renal function, and their chest X-rays were recorded to assess the degree of aortic arch calcification. The two variables Logistics regression was used to analyze independent risk factors of aortic arch calcification; Kaplan-Meier analysis was used to analyze the influence on prognosis of dialysis patients; and multivariate COX regression was employed to analyze independent risk factors of death in dialysis patients. Results Among the 177 selected cases of peritoneal dialysis patients, 66 cases (37.29%) presented with aortic arch calcification. Elevated serum phosphorus was an independent risk factor of aortic arch calcification (OR=54.69 ,95%CI:10.01-298.65, P<0.01). The probability of survival in patients with mild and moderate (severe) calcification of aortic arch was less than those without calcification. Moderate (severe) calcification of aortic arch was the independent risk factor of all-cause mortality and cardiovascular disease mortality, whose hazard ratios in patients with calcification were 3.779 times and 5.636 times of those in patients without calcification respectively. Conclusions Hyperphosphatemia is an independent risk factor promoting the development of calcification of aortic arch. The probability of survival in patients with mild and moderate (severe) calcification of aortic arch is less than those without calcification; moderate (severe) calcification of aortic arch is the independent risk factor of all-cause mortality and cardiovascular disease mortality.

OBJECTIVE:To study the improvement effect of lanthanum hydroxide on chronic renal failure (CRF) hyperphos-phatemia in rats. METHODS:CRF hyperphosphatemia rat model were induced and then randomly divided into model group,lan-thanum carbonate group [0.3 g/(kg·d)],calcium carbonate group [4.2 g/(kg·d)] and lanthanum hydroxide high-dose,medium-dose and low-dose groups [1.5,1,0.5 g/(kg·d)] with 10 rats in each group. They were given adenine 0.2 g/(kg·d)intragastrically in the morning,and then given relevant medicine intragastrically in the afternoon;a week later,they stopped taking adenine but con-tinued to take relevant medicine for 22 d. 10 normal rats were selected as normal control group. General examination was conduct-ed,and renal coefficient,serum contents of calcium,phosphorus,PTH,creatinine(Scr)and usea nitrogen(BUN)were detected after last medication as well as renal pathological change. RESULTS:Compared with normal control group,model group showed CRF sign,renal coefficient,the contents of phosphorus,PTH,Scr and BUN were increased,while the content of calcium was de-creased(P<0.01);renal section showed obvious pathological characteristics. Compared with model group,CRF sign of rats were improved in lanthanum carbonate group,calcium carbonate group and lanthanum hydroxide groups. The renal coefficient (except for lanthanum hydroxide high-dose group),serum contents of phosphorus(except for calcium carbonate group),PTH(except for lanthanum hydroxide low-dose group and calcium carbonate group),Scr(except for lanthanum hydroxide low-dose group and calci-um carbonate group)and BUN were all decreased,while serum content of calcium and calcium-phosphorucs product(only in calci-um carbonate group)was increased(P<0.05 or P<0.01). There was no statistical significance in other difference. The renal sec-tion pathological characteristics were improved. CONCLUSIONS:Lanthanum hydroxide can improve renal function and reduce the level of serum phosphorus in CRF hyperphosphatemia model rats.

OBJECTIVE: To explore effect of tobramycin (TOB) on healing of femoral fractures in rats. METHODS: Totally 32 male sprague-dawley (SD) rats were selected and randomly divided into sham group (group A), fracture group (group B), fracture with TOB group (group C) and fracture + TOB + IWR-1 group (group D), 8 rats in each group. Close femoral fracture model in rats were established in group B, C and D, group A was sham operation without otherwise process. Group D was intraperitoneal injected 100 μl (8 μM) of Wnt pathway inhibitor IWR-1-endo (IWR-1) before molding at 1 day. At 1 day after molding, 100 μl (100 μM) of TOB was intraperitoneally injected into group C and D at once a day for 7 days. At 7 weeks after modling, fracture healing of group B, C and D were observed by X-ray, Western blotting was appilied to detect alkaline phosphatase(ALP) and Runt related transcription factor 2 (RUNX2) and β-catenin of Wnt passway. RESULTS: X-ray results showed fracture line disappeared, callus formation and fracture healing well in group C compared with begning of molding; while a little fracture line, callus formation and fracture malunion in group B and d could be seen. Western blotting results showed ALP, RUNX2 and expression of β-catenin in group B, C and D were higher than that of group A ( CONCLUSION Tobramycin could promote osteoblast differentiation and fracture healing by stimulating Wnt / β-catenin signaling pathway, up regulating expression of ALP and RUNX2.
Alkaline Phosphatase ; Animals ; Cell Differentiation ; Core Binding Factor Alpha 1 Subunit/genetics* ; Femoral Fractures ; Fracture Healing ; Male ; Osteogenesis ; Rats ; Tobramycin ; Wnt Signaling Pathway ; beta Catenin/metabolism*

Background The in vitro culture of retinal vascular endothelial cells is the foundation of experimental study of retinal vascular disease. Shortage of human donor eyeballs is a main limiting for the laboratory work. The culture method of rat-derived vascular endothelial cells has been established. However, this method is not enough effective because of severer cellullar injury. Objective Present study was to establish a simple and high effective method for the culture of vascular endothelial cells in vitro. Methods The retinas from 5 SPF SD rats was digested by 0. 1% collagenase and cultured with explant culture method. 20% fetal bovine serum, vascular endothelial growth factor ( VEGF) , insulin-transferrin-selenium( ITS) were composed into the endothelial cell culture medium, and enough blowing was performed to get the cells and fragments from retinal tissue. The cellular suspension was prepared and cultured consequently on human fibronectin-coated culture flasks. Cultured vascular endothelial cells were identified by anti-von Willebrand staining factor. Results The cells emerged from the tissue mass,and cells and some tissue fragments attached to the wall after 24 hours of seeding. The cells grew to show the fusiform in 4 days and merged together in 5 to 6 days,and a cell monolayer was seen in the 14th day after culture. The endothelial cells showed the positive response for von Willebrand factor. After passage, the merging-growth statue of the cells was regained in 2 hours after culture. Conclusion Use of retinal pieces and collagenase-digestion can get the vascular endothelial cells with better activity in vitro. The culture method based on highly selective endothelial cell culture medium associated to FN adhesion-promoting is helpful for gaining the purified of endothelial cells.

Ewing's sarcoma/primitive neuroectodermal tumor (EWS/PNET) in the kidney is a rare but high-grade malignant tumor that affects predominantly elder children and adolescents.Patients mostly present with nonspecific symptoms such as abdominal pain and gross hematuria.Since EWS/PNET has a rapid clinical progression with early metastasis and death,it is essential to make an accurate and early diagnosis.Once diagnosed,multimodality treatment,including radical surgery combined with adjuvant chemotherapy,and radiotherapy if necessary,is recommended.Unfortunately,there are no characteristic signsthat have been described in ultrasonography or any other imaging modalities so far.The diagnosis of EWS/PNET is now based on a classical histological and immunohistochemical investigation complemented by a demonstration of specific chromosomal changes.Strong immunoreactivity to CD99 is ubiquitous,and t(11;22) translocation is seen in approximately 90％ of EWS/PNET.Herein,we report a patient with such condition.The patient was a young woman,and she presented with sudden right flank pain clinically.Ultrasonography revealed a large heterogeneous mass in the lower pole of her right kidney.The tumor compressed the renal pelvis and led to upper pole caliectasis.Color Doppler demonstrated blood flow with a pulsatile arterialized waveform within the mass.The patient received radical nephrectomy with right renal vein and vena cava thrombectomy.A search for other sites of tumor involvement yielded negative results.And six cycles of chemotherapy were sequentially performed.The diagnosis of EWS/PNET was confirmed based on primitive small round cell histology and characteristic immunohistochemical results.She was still alive with no evidence of recurrence five years after initial diagnosis.We would like to point out that ultrasound is still a useful method for initial assessment,and ultrasound-guided fine needle aspiration may play an important role in determining preoperative diagnosis.