Streptococcus pneumoniae(S.pn) is an opportunity pathogenic bacteria,environmental factors play a key role in the pathogenicity of S.pn. It is important to study virulent gene in vivo. The S.pn suicide plasmid containing gfp reporter was constructed by fusing the genes pneumolysin and gfp,in which gfp is an excellent molecule probe in vivo. The plasmid was integrated to No.22 S.pn by homologous recombination. The recombinant S.pn was gained and evaluated in aspects of fluorescence excitation, biological character and physio-activity. The results showed it is efficient and available to report the expression of virulent genes in vivo and in vitro, which will provide a new easy method for evaluating and screening the virulent genes of S.pn in vivo.

In this experiment six methods,calcium chloride(CaCl_(2)) precipitation,polyethylene glycol(PEG,pH7.0) precipitation,polyethylene glycol(PEG,pH11.5) precipitation,aluminum chloride(AlCl_(3)) precipitation,Amicon Utcra centrifugal filter devices and cellulose nitrate membrane were used to concentrate the vaccine poliovirus type 1(PV_(1)) added to water body;experimental conditions for concentration were selected and optimized.The results showed that two methods,CaCl_(2)and PEG(pH 7.0) precipitation were suitable for concentrating virus in large volumes of water while amicon utcra centrifugal filter devices for small ones.The virus recovery of the three methods reached a 100% rate.

Acupuncture Therapy ; Adult ; Chronic Disease ; therapy ; Humans ; Male ; Moxibustion ; Pelvic Pain ; therapy ; Prostatitis ; therapy ; Treatment Outcome ; Young Adult

Objective To summarize the experience of hybrid repair performed in high risk patients with dissection involving the aortic arch.Methods From Sep.2007 to Mar.2015,hybrid repair was performed in 33 high risk patients with dissection involving the aortic arch including acute (n =8),subacute (n =15),or chronic (n =10) cases.Descripitive statistics were computed for continuous and categorical variables.Results There were 22 male and 11 female patients with a mean age of(69 ± 10) years,and ASA Physical Status Ⅲ-Ⅳ.Simultaneous (n =27) and staged (n =5,mean interval 5.0 ± 1.3 days)endovascular repair were performed via femoral artery.The technical success rate was 100％.The average hospital stay was (16 ±6) days.One case died of cerebral infraction.There were two with strokes,one with pneumonia and two with renal failure as complications.Median follow-up was 47 months (3-66 months).There were four deaths with two were related to aortic artery.Endoleak was found in 3 during follow-up.One type Ⅰ endoleak was cured after remedy hybrid repair.Conclusions Hybrid repair performed in patients at high risk with dissection involving the aortic arch is less invasive with favorable medium and long-term outcomes.

Background Glaucoma is one of the leading causes of blindness worldwide;while the different types of glaucoma is vary from different region. Objective Present study was to survey the prevalence and types of glaucoma among peasants of Uigur adults in Kuche county and offer the basis for the prevent and treatment of glaucoma in Uigur nationanlity. Methods 4191 Uigur peasants aged 40 years or above were collected in Kuche county for the survey of prevalence and types of glaucoma by randomized cluster sampling in March and April of 2009. The subjects were grouped into 40-,50-,60- and ≥70 years groups according to the distribution of age. The disease history of glaucoma, regular eye examination, funds examination and measurement of the anterior chamber depth, gonioscope were performed in the all subjects. Darkroom prone test and mydriasis test were carried out in suspicious glaucomous patients. The depth of periphery anterior chamber was assessed based on van Herick' s criteria, and the width of chamber angle was graded based on Scheie' s method. The standardized training was performed. This survey approved by Xinjiang Medical Ethics Committee, all subjects signed the informed consent before the examination. Results 4191 of 4873 subjects finished all the examinations with the response rate 86%. All the subjects showed a good compliance. The prevalence of glaucoma was 3. 79% , and the prevalence of primary angle-closed glaucoma(PACG) .primary angle-open glaucoma (PAOG) and secondary glaucoma was 2.22% ,0.26% and 1. 31% respectively, showing a significant difference among the different types of glaucoma( P<0. 05). The prevalence of glaucoma was elevated with aging (χ2 - 116. 69 ,P<0. 05) and presented a high rate in male subjects compared with female ones(χ2 = 7. 34, P<0. 05 ). Bilateral blindness was found in 19.75% glaucoma peasants, in which 25.3% glaucoma peasants received anti-glaucoma surgery. The distribution of visual acuity of patients was of significant difference among different age groups(χ2 = 37. 69 ,P<0. 05 ) . Conclusions The prevalence of the glaucoma among Uigur peasants in Kuche county was higher than most area no matter inland or overseas. PACG still is the common type in those people.

BACKGROUND: The technology of biodegradable materials covering growth factors can be used to make sustained-release microspheres, which provides the feasibility for the efficient utilization of growth factors. OBJECTIVE: To prepare nano/micron-sized spheres using recombinant human bone morphogenetic protein 2/poly(lactic-co-glycolic acid) (rhBMP-2/PLGA)copolymer and to compare their release behaviors by in vivo and in vitro release experiments. METHODS: The rhBMP-2/PLGA nano/micron-sized spheres were prepared by emulsion solvent evaporation method to control the rate of pulp mixing.(1)In vitro release experiment:Prepared nano/micron-sized spheres were dissolved in PBS for 70 days,and then ELISA method was used to detect the rhBMP-2 concentration in the supernatant at different time.(2)In vivo release experiment:Forty-four New Zealand rabbits were divided into two groups, and rhBMP-2/PLGA nano/micron-sized spheres were respectively implanted into trochanteric defects. The concentration of rhBMP-2 in the defect site was detected by ELISA within 70 days after implantation. RESULTS AND CONCLUSION:In vitro sustained release experiment:There was a sudden release of nanospheres in the former 3 days,and the cumulative release nearly reached 41%, followed by a steady and slow release, and then the cumulative release was up to approximately 83% at 70 days. The initial release of micron-sized spheres was less than that of nanospheres, and the cumulative release was about 20% within the former 3 days and reached to 70% at 70 days.In vivo sustained release test:There was a sudden release of the nanospheres,the cumulative release was nearly 35%, followed by a steady and slow release, and then the cumulative release was up to approximately 72% at 70 days. The initial release of micron-sized spheres was less than that of nanospheres, and the cumulative release was about 21% in the former 3 days and increased to about 63% at 70 days.In both in vivo and in vitro release experiments,the release duration of micron-sized spheres was longer than that of nanospheres in the former 3 days. To conclude, the release time of rhBMP-2/PLGA micron-sized spheres fulfills the need of bone growth cycle, therefore, rhBMP-2/PLGA micron-sized spheres are more favorable than rhBMP-2/PLGA nanospheres for bone defect repair in clinical practice.

To analyze the characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy (DN) and its compliance with traditional Chinese medicine (TCM)symptom, for the sake of providing a basis for clarifying the rules of TCM syndrome differentiation in DN. Adopting the traditional epidemiological retrospective method, thirty-eight TCM syndromes and urinary protein with medium or low molecular weight, as well as urinary enzyme, including 24 h urinary protein (Upro), urinary albumin( UAlb), urinary retinal binding protein( URBP), urinary cystatin C (UCysC), urinary N-acetyl-beta-D-glucosaminidase (UNAG), were collected from 108 patients with stage III DN, and a multiple factor regression analysis between them was conducted. As the results, the levels of Upro, UAlb, URBP, UCysC, and UNAG were increased in 108 patients with stage III DN. Qi-Yin deficiency type was the major type. The level of UAlb in patients with Qi-Yin deficiency type was significantly higher than those without Qi-Yin deficiency type (P < 0.05). The elevation of Upro with the factors as swift digestion with rapid hungering, lassitude and lack of strength, weakness of waist and knees was complied, the elevation of UA1b with the factors as dry mouth with desire to drink, the elevation of URBP with the factors as numbness of extremities, shortness of breath, the elevation of UCysC with the factors as clear urine in large amounts, and the elevation of UNAG with the factors as frequent micturition, were complied respectively. In conclusion, for 108 stage III DN patients. The increase in urinary protein spectrum including UAlb, URBP, UCysC, and UNAG is the major characteristic. Shen and Pi are the major organs related to the appearance of urinary protein; Pi-Shen deficiency is the basic pathogenesis. The level of UAlb is taken as one of the objective syndrome factors for Qi-Yin deficiency type. The levels of UNAG and UCysC are possibly the objective syndrome factors for Shen-Qi deficiency type.
Diabetic Nephropathies ; complications ; diagnosis ; urine ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Proteinuria ; complications ; urine ; Qi ; Regression Analysis ; Yin-Yang

Objective To explore the influence of mononuclear cells (MNC), CD34+ cells, CD3+ , CD3+ CD4+ , CD3+ CD8+ , CD4+ CD25+ T cells, CD3- CD16+ CD56+ natural killer cells (NKs), and dendritic cells (DCs) doses in graft on acute graft-versus-host disease (aGVHD) following HLA-identical sibling allogeneic peripheral blood hematopoietic stem cell transplantation (allo-PBSCT).Methods Sixty-five patients receiving HLA-identical sibling allo-PBSCT were studied.The number of CD34+, CD3+, CD3+ CD4+, and CD3+ CD8+ T cells in the graft was counted by fluorescence-activated cell sorting (FACS).The number of CD4+ CD25+ T cells, CD3 CD16+ CD56+ NKs, and DCs in the graft was also measured by FACS in 31 patients among above-mentioned 65 patients.The doses of each kind of cells in the graft were calculated according to per kilogram of recipients body weight.The patients were divided into high or low dose groups according to whether or not more than or equal to median of MNC, CD34+, CD3+, CD3+ CD4+, CD3+CD8+, CD4+ CD25+, CD3 CD16+ CD56+ or DC cell doses, respectively.Acute GVHD was analyzed between two groups.Results The frequency of the cumulative incidence of grade Ⅱ～Ⅳ aGVHD was increased in CD3+ CD4+ and CD3+ CD8+ T cells high dose groups as compared with correspondingly low dose groups, but the difference had no statistically significant difference (P = 0.089 and 0.098, respectively).Recipients in CD4 + CD25 + T cells high dose group had significantly reduced cumulative incidence of grade Ⅲ～Ⅳ aGVHD as compared with those in correspondingly low dose group (P< 0.05).The cumulative incidence of total aGVHD was significantly higher in DC1 high dose group than in correspondingly low dose group (P<0.05) and the cumulative incidence of grade Ⅱ～Ⅳ aGVHD was also higher in high dose group, but the difference had no statistically significant difference (P = 0.069).There was no significant difference in cumulative incidence of total and grade Ⅱ～Ⅳ aGVHD between MNC, CD34+ , CD3+, NK or DC2 high dose groups and correspondingly low dose groups (P>0.05, respectively).Conclusion Recipients in DC1 high dose group have significantly increased cumulative incidence of total aGVHD, but those in CD4+ CD25+ T cells high dose group have significantly reduced cumulative incidence of grade Ⅲ～Ⅳ aGVHD.

In clinic, some urinary protein makers can dynamically and noninvasively reflect the degree of renal tubular injury in patients with diabetic nephropathy (DN). These urinary biomarkers of tubular damage are broadly divided into two categories. One is newfound, including kidney injury molecule-1 (Kim-1), neutrophil getatinase-associated lipocalin (NGAL), liver-type fatty acid-binding protein (L-FABP) and cystatin C (CysC); the other one is classical, including beta2 microglobulin (beta2-MG), retinal binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (NAG). It is reported that, the increases in urinary protein markers are not only closely related to the damage of tubular epithelial cells in DN patients, but also can be ameliorated by the treatment with Chinese herbal compound preparations or Chinese herbal medicine. Recently, although urinary proteomics are used in the protein separation and identification, the traditional associated detection of urinary protein markers is more practical in clinic. At present, it is possible that the associated detection of urinary biomarkers of glomerular and tubular damages may be a feasible measure to reveal the clinical significance of urinary protein markers in DN patients and the interventional effects of Chinese herbal medicine.
Biomarkers ; urine ; Diabetic Nephropathies ; complications ; drug therapy ; urine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; Proteinuria ; complications

Background The proliferation and migration of vascular endothelial cells is a primary link during angiogenesis.Studies showed that heat shock protein B6 (HspB6) promotes the secretion of multiple angiogenesis-related factors and therefore leads to neovascularization.Understanding the effects of neutralizing HspB6 antibody on the biological behavior of human choroidal vascular endothelial cells has an important significance in the target treatment of choroidal neovacularization diseases.Objective This study was to address the role and mechanism of neutralizing HspB6 antibody in tube formation of human choroidal vascular endothelial cells.Methods Human choroidal vascular endothelial cell line was normally cultured and harvested for total RNA extraction.Expressions of HspB6 mRNA and protein in human choroidal vascular endothelial cells were detected by reverse transcription PCR (RT-PCR) and flow cytometry (FCM).The cells were seeded on 96-well plate covered with matrigel at the density of 2×104/hole.Then the neutralizing HspB6 antibody at the concentration of 100 μg/Land 500 μg/L was added into the medium respectively,and the control cells were set without the addition of HspB6 antibody.The number of capillary tubes was calculated 12 hours after culture by three-dimensional matrigel assay.In addition,0,50,100,500 μg/L of neutralizing HspB6 antibody were added into the cell medium separately for 24hours,cell counting kit-8 (CCK-8) method was employed to assay the inhibitory rate(IR) of the cells.Transwell test was used to count the cell number across chamber membrane for the evaluation of migration ability of the cells.The apoptosis of the cells was assayed by FCM.Results Both HspB6 mRNA and protein were expressed on human choroidal vascular endothelial cells.The number of capillary tube formation of human choroidal vascular endothelial cells was (67.25±5.75),(60.39±6.41) and (39.76±10.73) /field in the 0,100 and 500 μg/L neutralizing HspB6 antibody groups,with significant difference among them (F =10.210,P =0.012),and the tube number was significantly less in the 500 μg/L neutralizing HspB6 antibody group compared with 0 μg/L neutralizing HspB6 group (P =0.005).The IR of neutralizing HspB6 antibody to the cellular proliferation and migration was enhanced with the increases of concentration and time lapse(Fconcentration =7.485,P =0.002 ; Ftime =16.684,P =0.001).The number of the cells through Transwell chamber membrane was 14.0 ± 2.5,11.1 ± 0.8,6.6 ± 0.1,6.7 ± 0.2 in the 0,50,100,500 μg/L neutralizing HspB6 antibody group respectively,and that in the 100 μg/L and 500 μg/L neutralizing HspB6 antibody group was lessened in comparison with the 0 μg/L neutralizing HspB6 antibody group(both at P=0.000).The apoptosis rate of the cells was (22.73 ± 2.53)％ in the neutralizing HspB6 antibody group,which was significantly lower than (13.33±2.08) ％ of the control group (t=4.967,P=0.008).Conclusions Neutralizing HspB6 antibody inhibits capillary tube formation of human choroidal endothelial cells in vitro in dose-and timedependent manner,probably through suppressing the proliferation and migration and promoting the apoptosis of choroidal endothelial cells.