Background Studieshowed thaDelta-like ligand 4 (Dll4) participatein the deveopmenof retinal celland angiogenesis.The Dll4-Notch pathway and vasculaendothelial growth facto(VEGF) are thoughto be critical mediatorof neovascularization undehypoxiconditions.The relationship between Dll4 and VEGF inovery cleaand furtheresearch ineeded.Objective Thistudy wato observe the inhibition of Dll4 on experimental retinal neovascularization and VEGF expression.MethodThe retinal neovascularization animal model wainduced by oxygen-induced retinopathy (OIR) in 5-day-old SPF SD ratby rearing the new postnatal ratwith the motherattogethein closed box with oxygen level a(80±2) ％ till 12-day-old.The ratwere then raised in normal aifo5 days.Aftethat,2.5μl (0.5 μg) of Dll4 monoclonal antibody wainjected into the mid-vitreoucavity in the righeye(Dll4 injected group) and PBwaused in the same way in the fellow eye(PBcontrol group) in the 12-day-old rats.Retinawere isolated in the 17-day-old rats,and retinal vasculamorphology waexamined by adenosine diphosphatease (ADPase) staining of retinal flatmounts,and the endotheliocyte nuclei above the internal limiting membrane were counted in the retinal tissue-slices.Reverse transcription PC(RT-PCR) waused to detecthe mRNexpression level of Dll4,VEGF,VEGF receptor-1 (VEGFR-1),VEGFR-2 and neuropilin-1 mRNin the retinas.Statistical analysiwaperformed by the paired t-test.The care and use of the animalcomplied with the Guidance Suggestion issued by the Ministry of Science and Technology of Chinin 2006.ResultThe Dll4 mRNexpression in the retin(Dll4 mRNA/β-actin mRNA) wa0.22± 0.06 and 0.98 ± 0.13 in the Dll4 injected group and the PBcontrol group,respectively,with statistically significandifference (=21.839,P =0.000).No significandifferencewere found in the expression of the VEGF mRNA,VEGFR-1 mRNand VEGFR-2 mRNin the retinabetween the two group(t=0.463,P=0.649;=1.687,P=0.109;=-1.674,P=0.111).Compared with the PBcontrol group,the expression of neuropilin-1 mRNwasignificantly elevated in the Dll4-injected group (0.73±0.08 vs.0.64±0.07) (t=-2.677,P=0.015).ADPase staining showed thathere were much more new blood vesselin the Dll4 injected group than those of the PBcontrol group.The numbeof nuclei structurally adjacento the vitreal side of the internal limiting membrane wa(63.6± 11.6)/slide in the Dll4 injected group,which wamore than thaof the PBcontrol group a(35.1±5.2)/slide (=-7.879,P =0.000).ConclusionDll4 playan essential role in the procesof pathological angiogenesiin the retina.Dll4 ithoughto be feedback regulatoof VEGFR,which participatein the procesof restraining pathological vasculogenesis.

OBJECTIVETo explore the clinical application of sural neurovascular pedicle fasciocutaneous reversed island flap to repair soft-tissue defection of the ankle and foot.

METHODSFrom Sep. 1994 to Oct. 2004,29 patients with soft-tissue defects in the ankle and foot were repaired by use of sural neurovascular fasciocutaneous reversed island flap, including 15 cases of traumatic defects, 11 cases of burns and 3 cases of chronic ulcer. The flap area ranged from 5 cm x 7 cm to 12 cm x 20 cm, and the length of pedicle from 5 cm to 12 cm.

RESULTSThe flaps survived totally in 27 cases, the distal necrosed partially and secondary free-skin grafting were further conducted in 2 cases. Twenty-one cases were followed-up for 3 to 60 months,the circulation, color and texture of the flaps were excellent and 2-point discrimination was 10 - 15 mm. The appearance and function of ankle joints were good.

CONCLUSIONThis flap has sufficient blood supply and a high survival rate; It is convenient in design, dissection and without sacrifice of major arteries. So, it is an effective method for the reconstruction of soft-tissue defects in ankle and foot.

Adolescent ; Adult ; Ankle Injuries ; surgery ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Middle Aged ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Sural Nerve ; blood supply ; Surgical Flaps ; blood supply ; Young Adult

OBJECTIVETo explore the effect of transforming growth factor alpha (TGFalpha) on the expression of cyclin E and D1 in gastric carcinoma cells.

METHODSHuman gastric adenocarcinoma SGC7901 cells were cultured routinely and synchronized at G(0)/G(1) phase in serum-free RPMI-1640. The percentage of the cells at G(0)/G(1) phase was detected by propidium iodide staining and flow cytometry (FCM), and the synchronized cells were cultured in RPMI-1640 supplemented with 2.5% calf serum and treated with 10, 30, and 50 microg/L TGFalpha for 5 h. The expression of cyclin E and D1 in SGC7901 cells was detected by immunofluorescent staining and FCM.

RESULTSThe percentage of the cells at G(0)/G(1) phase increased from 54% in routine culture to 72% in the serum-free RPMI-1640 culture. TGFalpha treatment of the cells synchronized at G(0)/G(1) phase induced significant increment of cyclin E and D1 expressions (P<0.001), and at the dose of TGFalpha of 50 microg/L, their expressions increased by 25.18% and 27.52%, respectively (P<0.001).

CONCLUSIONTGFalpha can increase the expression of cyclin E and D1 in gastric carcinoma cells to promote their cell cycle progress.

Adenocarcinoma ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cyclin D1 ; biosynthesis ; Cyclin E ; biosynthesis ; Flow Cytometry ; Fluorescent Antibody Technique ; Humans ; Stomach Neoplasms ; metabolism ; pathology ; Transforming Growth Factor alpha ; pharmacology

OBJECTIVETo evaluate the clinical practice of laparoscopic rectopexy in the treatment of severe rectal prolapse.

METHODSFrom March 1998 to February 2007, 4 cases of complete rectal prolapse, including 1 male and 3 female,ranged 21-82 years old, were treated by laparoscopic rectopexy. In one case, the posterior wall of rectum was freed and elevated, and pre-rectal introcession was closed by silk suture, then the posterior wall was suspended and fixed on sacral promontory fascia, finally the sigmoid colon was fixed by sutures on the fascia of left psoas major. In other three cases, insertion of mesh was performed. Rectum was freed and elevated to the level of levalor ani. A sheet of T-shape polypropylene mesh was placed posterior to the rectum, whose lower margin was at the level of levator ani and wrapped around the rectum covering except the anterior wall. The free margin of the mesh was sutured on the muscular layer of rectum, then the mesh was put posterior to the rectum and fixed on the sacral promontory fascia by clipping to repair hernia. After that, the pelvic peritoneum was closed, and finally the sigmoid colon was fixed by sutures on the fascia of left psoas major.

RESULTSFour operation procedures were completed successfully. There was no conversion operation. The time was consumed 92.5 (80-100) min, and the bleeding amount was 6.5 (5-10) ml. No post-operative complications were found. Urine incontinence and encopresis were relieved. No recurrence and constipation was found after 2 months to 3 years follow up postoperatively.

CONCLUSIONLaparoscopic rectopexy is a safe, workable and effective procedure, which can reduce operative trauma and shorten hospitalization time.

Aged ; Aged, 80 and over ; Female ; Humans ; Laparoscopy ; Male ; Rectal Prolapse ; surgery ; Rectum ; surgery ; Young Adult

OBJECTIVEX-linked inhibitor of apoptosis protein (XIAP) To gastrointestinal (GI) investigate the expression of XAF1 and heat-shock transcription factors 1 èHSF1éand their relationship in human gastrointestinal cancers.

METHODSImmunoblotting was used to analyze the expression of HSF1 and XAF1 in either gastric or colon cancer tissue and GI cancer cell line. Transient expression of the HSF1-containing vector in GI cell lines and RNA interference were used to up/down-regulae the expression of the HSF1, and the subsequent expression of XAF1 was measured.

RESULTSThe expression of HSF1 was higher in GI cancers than in normal tissues. The expression of XAF1 and HSF1 was negatively correlated in GI cancer cell lines. Stress stimuli up-regulated the expression of HSF1 while the alteration of XAF1 expression was negatively correlated with HSF1 expression.

CONCLUSIONThe high expression of HSF1 in GI cancers is associated with suppressed expression of XAF1, which can be one of the mechanisms for low-expression of XAF1 and insufficient apoptosis in GI cancers.

Animals ; Base Sequence ; Cell Line, Tumor ; DNA-Binding Proteins ; genetics ; Gastrointestinal Neoplasms ; genetics ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Heat Shock Transcription Factors ; Humans ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; genetics ; Oxidative Stress ; genetics ; Temperature ; Transcription Factors ; genetics

OBJECTIVETo investigate the expressions of X-linked inhibitor of apoptosis protein (XIAP)-associated factor-1 (XAFI) and heat-shock transcription factor 1 (HSF1) and their relationship in human gastrointestinal cancers.

METHODSImmunoblotting was used to analyze the expressions of HSF1 and XAF1 in gastric and colon cancer tissues and in gastrointestinal cancer cells. The gastrointestinal cancer cells were tranfected with a eukaryotic expression vector containing HSF1 gene fragment or subjected to RNA interference to induce up- or down-regulation of HSF1 expression, and the consequence changes in XAF1 expression in the cells was measured. XAF1 expression was also assayed in the cells after stress stimulation for HSF1 expression.

RESULTSThe expression of HSF1 was higher in gastrointestinal cancer tissues than in normal tissues. The expression of XAF1 and HSF1 was inversely correlated in the cancer cell lines, and stress stimuli of the cells up-regulated the expression of HSF1 but down-regulated XAF1 expression.

CONCLUSIONHSF1 expression is increased in gastrointestinal cancer tissues to result in suppressed expression of XAF1, which may be one of the reasons for the low expression of XAF1 in association with the defect of the apoptosis mechanisms in the cancer cells

Cell Line, Tumor ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; DNA-Binding Proteins ; biosynthesis ; genetics ; Heat Shock Transcription Factors ; Humans ; Immunoblotting ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; biosynthesis ; genetics ; RNA Interference ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transcription Factors ; biosynthesis ; genetics ; Transfection

OBJECTIVETo investigate the feasibility and safety of endoscopic thyroidectomy via the areola of breast approach.

METHODSBetween April 2005 to September 2008, endoscopic thyroidectomy via the areola of breast approach was performed in 28 female patients. Of the patients, 25 cases presented with nodular goiter, 2 cases with Grave's disease and 1 case with minimum papillary carcinoma. The average age was 22.5 years (range, 18-38 years). A 10 mm trocar was placed on the medial border of the areola of the right breast for the video-endoscopy and removing specimens, and a 5 mm trocar was placed on the lateral border of the areola of the same breast as the assisted operation hole. Another 5 mm trocar was placed on the medial border of the areola of left breast as the main operation hole. The operation data was recorded and analyzed.

RESULTSAll the 28 operations were successful. The procedures included one lobe total thyroidectomy in 5 cases, one lobe subtotal thyroidectomy in 15 cases, subtotal thyroidectomy in 3 cases, one lobe near total thyroidectomy + the other lobe subtotal thyroidectomy in 4 cases, and one lobe total thyroidectomy + the central group lymph node resection + the other lobe subtotal thyroidectomy in 1 case. The average operation time was 60.7 minutes (range, 40-125 minutes), the average operation blood loss was 5.8 ml (range, 2-15 ml), the average length of post-operative hospital stay was 3.1 days (range, 2-5 days). No adverse effects was found after the operation, such as damage to the parathyroid gland and the laryngeal nerve. The patients were followed-up for 1 to 40 months with satisfactory results. All 28 patients were satisfied with the cosmetic effects of the operation.

CONCLUSIONSEndoscopic thyroidectomy via the areola of breast approach produces an outstanding cosmetic effect, it is safe and feasible.

Adolescent ; Adult ; Breast ; surgery ; Endoscopy ; methods ; Female ; Follow-Up Studies ; Humans ; Thyroid Diseases ; surgery ; Thyroidectomy ; methods ; Treatment Outcome ; Young Adult

The immune checkpoint programmed cell death-ligand 1(PD-L1)-mediated immunosuppression is among the important features of tumor. PD-L1, an immunosuppressant, can induce T cell failure by binding to programmed cell death-1(PD-1). Thus, the key to restoring the function of T cells is inhibiting the expression of PD-L1. The Chinese medicinal Atractylodis Macrocephalae Rhizoma(AMR) has the anti-tumor, anti-inflammatory, antioxidant, and hypoglycemic activities, and the polysaccharide in AMR(PAMR) plays a crucial role in immunoregulation, but the influence on the immune checkpoints which are closely related to immunosuppression has not been reported. MicroRNA-34 a(miR-34 a) expression in esophageal carcinoma tissue is significantly lower than that in normal tissue. This study aims to investigate the inhibitory effect of PAMR on esophageal carcinoma cells, and the relationship between its inhibitory effect on PD-L1 expression and miR-34 a, which is expected to clarify the anti-tumor mechanism of PAMR. Firstly, different human esophageal carcinoma cell lines(EC9706, EC-1, TE-1, EC109 cells) were screend out, and expression of PD-L1 was determined. Then, EC109 cells, with high expression of PD-L1, were selected for further experiment. The result showed that PAMR suppressed EC109 cell growth. According to the real-time quantitative PCR(qPCR) and Western blot, it significantly suppressed the mRNA and protein expression of PD-L1, while promoting the expression of tumor suppressor miR-34 a. The confocal microscopy and luci-ferase assay proved that PAMR alleviated the inhibitory effect of PD-L1 while blocked miR-34 a. Additionally, the expression of PD-L1 was controlled by miR-34 a, and the combination of miR-34 a inhibitor with high-dose PAMR reversed the inhibitory effect of PAMR on PD-L1 protein expression. Thus, the PAMR may inhibit PD-L1 by increasing the expression of miR-34 a and regulating its downstream target genes. In conclusion, PAMR inhibits the expression of PD-L1 mainly by inducing miR-34 a.
B7-H1 Antigen/pharmacology* ; Carcinoma ; Cell Proliferation ; Humans ; MicroRNAs/metabolism* ; Polysaccharides/pharmacology*

This work is to establish the fingerprint of Astragalus membranaceus var. mongholicus by HPLC-ELSD method, and to analyze the simulated wildness degree of A. membranaceus var. mongholicus in the genuine region of Inner Mongolia, Ningxia and Gansu. Compared with wild A. membranaceus var. mongholicus, the quality differences of A. membranaceus var. mongholicus in the genuine region were analyzed by identification of chromatographic peaks and similarity evaluation, cluster analysis(CA), principal components analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). HPLC fingerprints of A. membranaceus var. mongholicus in different genuine regions are established. The qualitative analysis of mass spectrometry identified 18 components. The similarity evaluation shows that the similarity of 32 batches of A. membranaceus var. mongholicus samples was 0.688-0.993. Among them, the similarity of samples in Shanxi, Inner Mongolia, Ningxia is 0.688-0.993, 0.835-0.989, 0.934-0.988, respectively and the similarity of samples in Gansu is 0.729-0.876 except No. 25 sample. The results of CA show that the samples of A. membranaceus var. mongholicus can be grouped into four categories according to the production area except the No. 11 and No. 25 samples. The results of PCA indicate that 32 batches of A. membranaceus var. mongholicus samples can be clustered according to quality and origin, and the quality of A. membranaceus var. mongholicus in Inner Mongolia is the closest to the wild breed. The results of OPLS-DA indicate that there are six components that can distinguish the wild and domestic A. membranaceus var. mongholicus, which are malonylastragaloside Ⅰ, astragaloside Ⅰ, calycosin-7-O-β-D-glycoside-6″-O-malonate, calycosin-7-O-β-D-glycoside, formononetin-7-O-β-D-glycoside-6″-O-malonate, and astrapterocarpan-3-O-β-D-glycoside-6″-O-malonate. The established method can be used to analyze differences between A. membranaceus var. mongholicus origin and planting environment, and can provide references for the protection and replacement of wild A. membranaceus var. mongholicus resources, and the cultivation, processing and production of A. membranaceus var. mongholicus.
Astragalus propinquus ; China

In recent years,the development and application of classical famous prescriptions have attracted much attention. However,the differences between ancient and modern conditions lead to difficulties in carrying out practical work. In this paper,with Houpu Wenzhong Decoction as an example,the key technologies of boiling granularity,water addition,boiling time and sample pretreatment methods were investigated on the basis of sufficient literature research. The experimental results showed that there was no significant difference in the concentration of index components between those with different granularity( 2 mm and 3-5 mm) and different decocting time( 30 min and 60 min),but the extraction rate of index components was relatively high when the granularity of powder was 2 mm and decocting time was 30 min. With the increase of water content,the concentration of index components and the extraction rate were increased in varying degrees. A certain proportion of methanol aqueous solution was used as the resolvent before content determination of the reference sample of Houpu Wenzhong Decoction,which could take into account both the spectral information of water-soluble components and fat-soluble components in the prescription,and help to display the overall information of the prescription' s chemical components more comprehensively. At the same time,the boiling and dispersing classical prescriptions in the Catalogue of Ancient Classical Prescriptions( the first batch) were collected and summarized in this study; the key influencing factors of decocting process were analyzed from different angles,and preliminary research suggestions were put forward,so as to provide a certain direction and reference for the establishment of quality standard of Houpu Wenzhong Decoction,as well as for the development,research and clinical application of boiling and dispersing classical prescriptions.
Drugs, Chinese Herbal/standards* ; Powders ; Prescriptions