Background Alkali burn-induced corneal neovascularization(CNV) usually leads to blindness.Recently,study determined that vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) were the main regulating factors inducing angiogenesis,and canstatin proteins can inhibit the growth of VEGF and bFGF and thereby inhibit CNV growth.Objective The present study was to investigate the effect of recombinant canstatin protein on mouse CNV induced by alkali burn and its mechanism.Methods CNV models were induced in the right eyes of 40 female BALB/c mice by sticking the 2.0 mm filtering paper with 1 mol/L NaOH at the central cornea for 10 seconds.The animals then were randomized into two groups.Recombinant canstatin protein drops(5 mg/L) was topically administered 4 times daily in the mice of the experimental group,and normal saline solution was used at the same way in the control group.Corneas of the mice were examined under the slit lamp to calculate the CNV area 1 day,3,7,14 days after modeling.The mice were sacrificed at above time points and corneas were obtained.The expressions of VEGF protein and bFGF protein in cornea were detected by Western blot,and the results were analyzed by enhanced chemiluminescence(ECL).The use of the experimental animals complied with the Instructive Notions with Respect to Caring for Laboratory Animals by State Ministry of Science and Technology.Results In 3,7 and 14 days after establishment of models,the area of CNV was (1.98-0.31) mm2,(6.21 ±0.44) mm2 and (9.83±0.72) mm2 in the experimental group,and that in the control group was (2.92 ± 0.41) mm2,(8.04 ± 0.56) mm2 and (11.78 ±0.84) mm2,showing significant reduce in the mice treated with recombinant canstatin protein(t3d =4.332,P=0.005 ;t7 d =11.729,P =0.000 ;t14 d =14.562,P =0.000).Western blot analysis showed that there was significant increase in the gray scale of VEGF protein 1 day,3,7,14 days following alkali burn in the experimental group compared with the control group(t1 d =-3.980,P＜0.001 ;t3d =-10.020,P＜0.001 ;t7d =-4.355,P＜0.001 ;t14 d =-8.156,P＜0.001),and the gray scale of bFGF was significantly ascent at various time points in the the experimental in comparison with the control groups (t1 d =-3.488,P＜0.001 ; t3 d =-2.124,P =0.013; t7 d =-1.977,P =0.028; t14 d =-4.542,P＜0.001).Conclusions Topical application of recombinant canstatin protein drops inhibits CNV growth induced by alkali burn by down-regulating the expressions of VEGF and bFGF proteins.

Objective To identify and rename one strain stored in National Center for Medicine Culture Collections ( CMCC) and used for tracheitis vaccine production. Methods The test strain CMCC (B)29108 and the type strain DSM30007T were cultured on NA medium. Characteristics in morphology, physiology, biochemistry and fatty acid profile were compared between the two strains. Phylogenetic analysis was based on 16S rRNA and rpoB gene sequences, together with the DNA-DNA hybridization assay. Results A Comparative analysis of a partial sequence of the 16S rRNA gene sequence revealed that the CMCC( B) 29108 strain was closed to the Acinetobacter species and showed the highest similarity with the type strain Acinetobacter baumannii DSM30007T. Moreover, the CMCC(B)29108 strain was highly similar to type strain DSM30007T in morphology, physiology, biochemistry and fatty acid profile. On the phylogenetic tree based on 16S rRNA and rpoB gene of all Acinetobacter members, the CMCC(B)29108 strain steadily clustered into one independent branch only with the DSM30007 T strain with a DNA-DNA hybridization value of 100%. Conclusion The CMCC(B)29108 strain that is one of the strains used for the production of tracheitis vac-cine should be assigned to the species of Acinetobacter baumannii based on its phenotypic, phylogenetic and chemotaxonomic characteristics.

Objective To investigate the relationship between apparent diffusion coefficient (ADC)value of rectal adenocarcinoma on DWI and its pathological grading.Methods The ADC values of 46 rectal adenocarcinomas were measured and compared with their histopathological grades.Results The 46 rectal adenocarcinomas included well differentiated adenocarcinomas in 14,moderate-ly differentiated ones in 20,and poorly differentiated ones in 12.The ADC values of well,moderately and poorly differentiated ade-nocarcinomas were (1.125±0.103)×10 -3 mm2/s,(1.030±0.098)×10 -3 mm2/s and (0.922±0.091)×10 -3 mm2/s,respective-ly,exhibiting a statistical difference (χ2 =1 7.35 1,P =0.000).Mann-Whitney U test showed that difference in ADC value between different histopathological grades was statistically significant.Conclusion ADC value of rectal adenocarcinoma can be used as a bio-marker for cell grading to guide treatment decision and prognosis assessment.

[Objective] To researchthe influence of endometrectomy and hysterectomy on the physiology and mentality of patients with refractory menorrhagia.[Method] This research selected 110 patients with refractory menorrhagia that accepted in our hospital from January,2015 to January,2017 as the object of study.Under the consent of patients,they were randomly divided into the endometrectomy group and hysterectomy group.Then the rate of Self-rating Anxiety Scale (SAS) and Self-rating Depression Scale (SDS),the recovery condition of colporrhagia and improvement of anemia of both groups were compared.[Result] Before treatment,the compare between both group's rate of SAS and SDS are not statistically significant (P＞0.05),after treatment,the SAS and SDS rate of endometrectomy group are respectively (45.31±3.58) and (46.31±3.46),which are both significantly lower than the hysterectomy group (P＜0.05);the endometrectomy group has a good rate of 94.55％ on the recovery condition of colporrhagia,which is significantly higher than the hysterectomy group's 81.81％ (P＜0.05);before surgery,the compare of hemoglobin quantity between two group are not statistically significant (P＞0.05);within 1,3 and 6 months after surgery,the hemoglobin level of endometrectomy group are significantly higher than the hysterectomy group (P＜0.05).[Conclusion] Both endometrectomy and hysterectomy can effectively treat patients with refractory menorrhagia,but endometrectomy has less influence on patient's physiology and mentality,which can facilitate patient's recovery and could be widely applied on clinical treatment.

Objective To investigate the effects of pulsed electromagnetic fields(PEMFs)on proliferation and differentiation of endothelial progenitor cells(EPCs).Methods EPCs were isolated from rat bone marrow by density gradient centrifugation.EPCs were exposed to PEMFs from the 5th day to the end of culture.MTT was used to measure the proliferation of EPCs.The expression ofⅧ-related antigen and NOS_3 was evaluated by flow cytometry. Results Compared with the control,the proliferating ability of EPCs exposed to PEMFs was stronger;the number ofⅧ-related antigen and NOS_3 positive cells increased significantly in EPCs exposed in PEMFs.Conclusion PEMFs promotes the proliferation and differentiation of rat bone marrow EPCs.

Objective To investigate the expression of exogenous gene transferred by piggyBac (PB) transposon in various gynecological malignant cell lines and reveal its potential application of gene therapy in gynecological cancer.Methods Amplified herpes simplex virus thymidine kinase (HSV-tk) gene coding region by PCR and integrated it into PB expression vector, PB[Act-RFP]DS, for reconstructing PB[Act-RFP, HSV-tk]DS (pPB/TK).By using different transfection reagents: FuGENE HD, jetPEI, lipofectamine 2000, pPB/TK together with helper plasmid Act-PBase were cotransfected into four mostly common gynecological malignant tumor cell lines HeLa, JEG-3, SKOV3 and HEC-1B.The mRFP1 report gene expressions was observed and detected by fluorescence microscope and flow cytometry to analyze transfection efficiency.The expressions of HSV-tk and mRFP1 gene were detected by reverse transcription PCR (RT-PCR).The cytotoxic effect of various concentration of pro-drug ganciclovir (GCV) for transfected cells was detected by methyl thiazole tetrazolium assay.The transfected cells were positive sorted by flow cytometry and limiting diluted to obtain the stable transfected cell line.The insertion sites of foreign gene tranferred by PB transposon in genome were analyzed by inverse PCR.Results (1) Double digests analysis and sequences test demonstrated that pPB/TK vector was reconstructed successfully.(2) Using three different transfective reagents, PB trausposon transferred HSV-tk gene and mRFP1 gene into HeLa, HEC-1 B, SKOV3 and JEG-3 cell efficiently, and the transfection efficiency of pPB/TK for the same cell was different by using different transfective reagents; in Hela cell, the transfection efficiency of FuGENE HD [(78.7 ± 9.2) %]was higher than that of lipofectamine 2000 [(54.1 ± 11.4) %]and jetPEI [(46.5 ± 7.4) %, all P < 0.05] ; using the same transfective reagent, the transfection efficiency of pPB/TK was also different on various cell lines, using FuGENE HD, the transfeetion efficiency of pPB/TK on HeLa, JEG-3 and SKOV3 cell was (78.7 ± 9.2) %, (74.4 ± 8.9) % and (83.2 ± 9.7) % respectively, which all were higher than that on HEC-1B [(39.5 ± 8.7) %, P < 0.05] .(3) RT-PCR showed that there were the mRNA expression of HSV-tk and mRFP1 in all cell lines.(4) 50% inhibitory concentration of GCV for transfected cells, HeLa, JEG-3, SKOV3 and HEC-1B, was 1.29, 3.35, 0.09 and 13.28 μg/ml respectively.Inhibitory effect of GCV (10 μg/ml) on SKOV3 transfected with pPB/TK was (86 ± 9) %, which was superior to that transfected with pORF-HSVtk alone [(52 ± 12)%, P < 0.05] .(5) The insertion sites of PB transposon in the target cells genome were located at TTAA sites, mRFP1 expression still could be detected in three months after transfected.Conclusions PB transposon could transfer exogenous gene into various gynecological malignant cells, which could integrated into genome and obtain a long-term and stable expression.It is expected that PB transposon may supply a more efficient and safer transgene technology platform for gene therapy in gynecological cancer.

This paper analyses the defects of bubble oxygen inhalators currently used, and investigates into their solutions for improvement.
Oxygen Inhalation Therapy ; instrumentation ; methods ; Oxygenators ; standards

OBJECTIVE:To provide references for clinicians in the choice of antihypertensive drugs.METHODS:The antihypertensive efficacy 6 classes(or 15 kinds) of antihypertensive drugs in a monotherapy in the treatment of 370 patients with essential hypertension(EH) were compared.RESULTS:The blood pressure was effectively lowered by all of the antihypertensive drugs except doxazosin,and doxazosin and torasemide showed inferior efficacy in the reduction of diastolic blood pressure(DBP).CONCLUSION:Calcium antagonists with prolonged action,angiotensin converting enzyme inhibitor(ACEI),angiotens-in receptor antagonist,and the monotherapy of Terazosin or Indapamide can all effectively bring down the blood pressure.It is suggested that low-dose doxazosin and torasemide should not be used alone in treating essential hypertension.

Objective To describe the skeletal CT imaging manifestations in patients with tuberous sclerosis complex(TSC),and to analyze their diagnostic value so as to establish an adequate skeletal change imaging data for the diagnosis of TSC.Methods Thirteen patients fulfilling TSC diagnostic criteria were examined with CT of the brain (n=13)and abdomen (n=7).Examinations from January,2004 to July, 2006 were retrospectively analyzed.Results There were three forms of lesions being demonstrated on CT: (1)Multiple sclerosing nodule (n=13):numerous,ovoid and circular,homogeneous,small and well- defined loci and symmetrical lesions were revealed in all cases in the central marrow portion of the bones, which could mimic blastic metastases.Follow-up CT imaging showed no change in both size and number. The lesions measured approximately 2-10mm.(2)Local sclerosing bone dysplasia with little bone expansion (n=7).Symmetrical and irregular density in the radix of the posterior arch of the vertebral body (n = 5 ).(3 )The spherical periosteal proliferation demonstrating as a cortex double line sign (n=2 ),and cortical thickening of metatarsals (n = 3 ).The appearance of the skeletal manifestation was as that in adulthood.Conclusion CT imaging of the skeletal system in TSC has some characteristics,by which the diagnosis of TSC could be made if combined with other main clinical diagnostic criteria. We suggest that those particular findings can be added as primary diagnostic features in the clinical diagnosis of TSC.

OBJECTIVETo establish a more stable method to isolate osteocytes in vitro, and then to find the differences with osteoblast biological characteristics.

METHODSOsteocytes and osteoblasts were isolated from the bone tissue of 3-day-old rats using sequential collagenase digestion. The cells were identified through cell morphology after 24 hours later. Alkaline phosphatase (ALP) kit was used to stain the first generation cells by Kaplow-way, the bone gla protein (BGP) of the cells were stained by immunocytochemitry. Measured ALP and computed its activity.

RESULTSOsteocytes and osteoblasts showed obviously differences in cell morphology. Osteocytes were star-shaped or dendrite-shaped within more dendrites, while osteoblasts were spindle-shaped with short dendrites. Osteocytes were negative for ALP, but osteoblasts were positive; Osteocytes were more positive for BGP, and osteoblasts were less positive. The secretion of ALP in osteocytes was lower than that of osteoblasts.

CONCLUSIONOsteocytes can be isolated and cultured in vitro. These characteristics of osteocytes are apparently difference with those of osteoblasts.

Alkaline Phosphatase ; analysis ; Animals ; Cell Separation ; methods ; Osteoblasts ; chemistry ; cytology ; Osteocalcin ; analysis ; Osteocytes ; chemistry ; cytology ; Rats ; Rats, Sprague-Dawley