Objective:To detect the degree of oxidative stress in the process when Porphyromonas gin-givalis ( P. gingivalis) stimulates human vascular endothelium, And to investigate the effect of peroxi-some proliferator-activated receptor(PPAR)γ on oxidative stress during this process. Methods:Human vascular endothelial cells ( HVECs) line EA. hy926 ( American Type Culture Collection ,United States) was cultured in high glucose Dulbecco' s modified eagle medium ( DMEM) . Four groups were designed:control group, P. gingivalis infected group, PPARγactivated group and PPARγblocked group. In con-trol group HVECs were cultured with only DMEM. In P. gingivalis infected group, HVECs were time-dependently stimulated by P. gingivalis W83 from 0 to 12 h. In PPARγ activated group or PPARγblocked group, PPARγ was pre-activated or blocked by a representative PPARγ agonist(15d-PGJ2 10μmol/L) or antagonist ( GW966210μmol/L) 30 minutes before the cells were stimulated by P. gingiva-lis. At 0, 0. 5, 1, 1. 5, 2, 4, 8, and 12 h, the culture medium was collected individually and centri-fuged, and the supernatant was stored for assay. Glutathione peroxidase (GSH-PX) and malondialdehyde( MDA) were analysed by enzyme-linked immunosorbent assay. Cellular reactive oxygen species ( ROS) were detected through 2',7'-dichlorofluorescin diacetate (DCFA-DA) fluorescent probe at various time points of the different groups. Results:In P. gingivalis infected group, the levels of GSH-PX [(5. 56 ± 0. 97) μmol/L] and MDA [(0. 84 ± 0. 18) nmol/L] were significantly higher than those in control group [GSH-PX(4. 71 ± 0. 64) μmol/L, MDA (0. 59 ± 0. 18) nmol/L)]. The levels of GSH-PX and MDA in PPARγactivated group [GSH-PX (5. 38 ± 0. 84) μmol/L, MDA (0. 84 ± 0. 22) nmol/L] and in PPARγblocked group [GSH-PX (5. 37 ± 0. 76) μmol/L, MDA (0. 85 ± 0. 14) nmol/L] were signi-ficantly higher than those in control group (P <0. 05). In the PPARγ activated group, the levels of GSH-PX at 0 . 5 and 8 h were significantly higher than those from 1 . 5 h to 4 h ( P<0 . 05 ) , while no difference was observed on the MDA levels at different time points. There was no significant difference at various time points for the levels of GSH-PX and MDA in PPARγ blocked group. The level of cellular ROS detected by DCFH-DA in P. gingivalis infected group was significantly higher than that in control group (10 108. 65 ± 1 805. 18 vs. 6 049. 06 ± 1 199. 19,P<0. 05). No difference was observed be-tween PPARγ activated group (7 120. 94 ± 1 447. 30) or PPARγblocked group (6 727. 35 ± 1 483. 68) and control group. Conclusion:Oxidative stress happens when P. gingivalis stimulates human vascular endothelium. PPARγ may involve in modulating oxidative stress during this process.

An extracellar fibrinolytic strain was isolated from fermented shrimp paste. In addition to general physiological and biochemical properties, the strain was identified by 16S rDNA sequence and systematic analysis. The results showed that 16S rDNA sequence of the strain had high similarity with AY601723 and AB195282, suggesting that the strain is a subspecies of Bacillus sp. It was named as Bacillus sp. nov. SK006 by CCTCC. The medium composition and fermentation conditions for fibrinolytic enzyme production were also optimized in the research.

Alpha-glycerophosphate oxidase (alpha-GPO) from Enterococcus casseli flavus was successfully isolated and purified by using polyethylene glycol (PEG)/(NH4)2SO4 aqueous two-phase system (ATPS). The results showed that the chosen PEG/(NH4)2SO4 ATPS could be affected by PEG molecular weight, pH, concentration of PEG and (NH4)2SO4, and inorganic salt as well as additional amount of crude enzyme. After evaluating these influencing factors, the final optimum purification strategy was formed by 16.5% (m/m) PEG2000, 13.2% (m/m) (NH4)2SO4, pH 7.5 and 30% (m/m) additive crude enzyme, respectively. The NaCl was a negative influencing factor which would lead to lower purification fold and activity recovery. These conditions eventually resulted in the activity recovery of 89% (m/m), distribution coefficient of 1.2 and purification fold of 7.0.
Ammonium Sulfate ; chemistry ; Glycerolphosphate Dehydrogenase ; chemistry ; isolation & purification ; Molecular Weight ; Polyethylene Glycols ; chemistry ; Water

Objective To probe into the application of PureceptionTM gradient centrifugation technique for sperm separation in artificial insemination by husband (AIH) .Methods Prepare the semen by PureceptionTM gradient centrifugation technique ,and used for 1360 AIH cycles in 672 infertile couples .Results After preparing the semen by PureceptionTM gradient centrifugation technique ,the sperm concentration was increased from (54 .86 ± 26 .03) × 106/mL to (63 .89 ± 34 .40) × 106/mL ,the rate of PR sperm was increased from (32 .47 ± 9 .31)% to (69 .78 ± 5 .10) ,the differences were both statistically significant (P<0 .05) .the recovery rate of PR sperm was (34 .08 ± 3 .95)% .672 infertile couples received 1360 AIH cycles ,and the clinical pregnance rate was 16 .62% (226/1360) .According to the sperm concentration before preparing the semen ,the cycles were divided into 3 groups :≥15 × 106/mL (GroupⅠ) ,(≥10 ,<15)× 106/mL (GroupⅡ ) and <10 × 106/mL (Group Ⅲ ) ,the clinical pregnancy rates were 17 .13% ,16 .05% and 4 .26% ,respectively ,Group Ⅲ was significantly lower than the other two groups (P<0 .05);According to the rate of PR sperm ,the cycles were divided into 3 groups:≥32% (Group A) ,(≥10 ,<32)% (Group B) and <10% (Group C) , the clinical pregnancy rates were 17 .53% ,16 .55% and 4 .65% ,respectively ,Group C was significantly lower than the other two groups (P<0 .05) .According to the number of PR sperm after preparing the semen ,the cycles were divided into 2 groups:≥10 × 106 and <10 × 106 ,the clinical pregnancy rates were 17 .33% and 10 .22% ,the difference was statistically significant (P<0 .05) . Conclusion PureceptionTM gradient centrifugation technique is a better way for sperm separation ,and the clinical pregnance rate is satisfactory when it is used in AIH .When the sperm concentration is ≥10 × 106/mL and the rate of PR sperm is ≥10% before pre‐paring the semen ,and the number of PR sperm is ≥10 × 106 after preparing the semen ,patients will get a more ideal pregnancy rate .

200 papers on nerve root type cervical spondylosis treated with Chinese medicine were retrieved and 38 papers with complete diagnostic criteria and medical statistics were included for study. The results showed acupuncture, massage, and herbal therapy were three common methods and have their own advantage, but systemic, standardized and normative treatment program was lack. In the meantime of treating nerve root type cervical spondylosis, prevention should also be paid attention. The treatment, prevention and exercise on the whole therapeutic idea should be established, which has far-reaching significance.

Objective To investigate the effect of neutrophil extracellular traps (NETs) on inflammation of rheumatoid arthritis (RA), especially angiogenesis. Methods The presence of NETs in synovial tissues of RA and osteoarthritis (OA) was observed by immunofluorescence assay. Neutrophils were isolated from peripheral blood of health volunteers. Neutrophils were cultured in vitro, the formation of NETs was observed. NETs were extracted as a stimulating agent. The effects of NETs on the proliferation of human umbilical vein endothelial cells (HUVECs) and synovial fibroblasts (RAFLS) were evaluated by MTT, and which were classified into two groups: HUVECs group and RAFLS group, with the following treatment: control and NETs (0.28 mg/L). Wound repair assay was employed to evaluate the effect on the cell migration stimulated with NETs. The experiment was divided into three groups:control, VEGF (40μg/L VEGF) and NETs (0.28 mg/L NETs). Results (1) Compared with OA, NETs were found more in the synovial tissue of RA. (2) NETs formation was induced by stimulator in vitro. The concentration of extracted NETs-DNA was 27.8 mg/L. (3) MTT assay showed that compared with the control groups, low concentration of NETs (0.28 mg/L) promoted the proliferation of HUVECs (0.499 ± 0.011 vs. 0.393 ± 0.009, P<0.05) and RAFLS (0.266 ± 0.007 vs. 0.192 ± 0.007, P<0.05). (4) It was showed that a significant wound closure induced by low concentration of NETs (0.28 mg/L) was found compared with control. Conclusion Our present study suggests that NETs are found more in the synovial tissue of RA, and low concentration of NETs can promote angiogenesis in RA.

ObjectiveTo identify the correlation factors for urosepsis shock after endourology surgery and analyze its clinical characteristics.MethodsFifty-eight patients with urosepsis without shock and 15 patients with shock were studied.Factors which may be correlated with the development of sepsis were analyzed including age,sex,ASA physical status,history of urinary tract infection,preoperative leukocyturia,urine bacteria culture,prophylactic antimicrobial treatment,pyonephrosis,type of surgery,duration of operation,blood leukocyte count and concentration of creactive protein at the end of surgery,the time when the signs of infection (shivering,agitation,nausea or vomiting) first appeared.Changes in BP in patients with urosepsis shock,scores for multiple organ dysfunction syndrome as well as treatment and prognosis were also recorded.ResultsFemale sex and early emergence of signs of infection after surgery were correlated with urosepsis shock.Abrupt hypotension was usually the first manifestation of urosepsis shook.The lowest systolic BP was positively correlated with the time when diagnosis of shock was made and negatively correlated with the length of ICU stay and the highest multiple organ dysfunction syndrome scores but not correlated with shock correction time.All patients with urosepsis shock recovered completely when discharged from hospital.ConclusionFemale sex and patients with early emergence of signs of infection are prone to develop urosepsis shock after endourology operation.The prognosis of urosepsis shock is good if diagnosed and treated in time.

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

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 compare the different effects of 1% diatrizoate meglumine,2.5% mannitol and water as oral contrasts in PET/CT scan in gastrointestinal tract delineation and 18F-fluorodeoxyglucose (FDG) uptake. Methods Sixty-one patients referred for PET/CT scan without gastrointestinal diseases were divided into three groups randomly ( random number method). One liter of 1% diatrizoate meglumine,2.5% mannitol,or water was orally taken by groups 1 (25 cases),2 (20 cases) and 3 ( 16 cases),respectively before scan. The scan was performed with GE Discovery LS PET/CT scanner in two-dimensional (2D) mode 50 min after 18F-FDG (5.55 MBq/kg) injection. Patients with abdominal lesions were excluded from this study. The degree of gastrointestinal filling and 18F-FDG uptake was evaluated by 3 nuclear medicine physicians using visual analysis according to a 4-grade classification method:none,mild,moderate,and high. Statistically analysis was performed by Kruskal-Wallis,Mann-Whitney and paired t tests.Results Both the differences of serum glucose and insulin levels were not significant before and after contrast taken in group 2. Group 2 had better gastrointestinal filling than that of group 1 and also better than group 3 except in rectum. The stomach,jejunum,ascending,and transverse colon were better filled in group 1 than in group 3. The degree of 18F-FDG uptake of group 3 was significantly higher than that of group 2 in stomach,jejunum and ileum (z= -3. 192,-3.290,-3.290,all P＜0.05),and was also significantly higher than that of group 1 (z = - 3. 603,P ＜ 0.05) in jejunum. The degree of 18 F-FDG uptake of group 3 was significantly lower than that of group 1 in ascending colon (z = - 2. 706,P ＜ 0. 05 ) and was significantly lower than that of group 1 and 2 in transverse and descending colon (z= - 3. 503,- 2.403,- 4.225,-4. 027,all P ＜0.05),and was also significantly lower than that of group 2 in rectum (z = -4. 128,P ＜0. 01 ). The maximum CT values in stomach,jejunum,ileum and ascending colon in group 1 were ( 132 ±23),(191 ±31),(313 ±47) and (374±53) HU,respectively,whose difference was significant (t = -7.088--1.781,all P ＜0. 01 ). Conclusion Oral iso-osmotic mannitol intake has better gastrointestinal filling and less physiological 18F-FDG uptake compared to diatrizoate meglumine and water.