Chemical disinfectants are generally used for virus inactivation and environment disinfection in biosafety laboratory, and the efficacy and evaluation of the disinfection are critical to ensure the laboratory biosafety.However, there is a current lack of applied standard to evaluate the virucidal efficacy of chemical disinfectants in our country.In this paper, a European Union standard“Method and Requirements of Virucidal Quantitative Suspension Test Method for Chemical Disinfectants Used in Human Medicine” was analyzed and a standard transformation scheme has been proposed.It is suggested that the model viruses should be increased from 3 to 6, including the surrogate viruses to substitute highly pathogenic viruses, and that the method to remove the residual chemical disinfectant and the calculation of 95%confidence interval should be incorporated into the standard.The suggestion will improve the scientific and operational standards related to disinfection and sterilization in biosafety laboratory in China.

Objective To analyze the effects of minimally invasive intenvention on acute suppurative cholangitis(ASC).Methods The clinical data of 28 patients with ASC and performed with endoscopic therapy were analyzed.Results 5 patients with a single stone incarcerated in the duodenal nipple were performed with ERCP and needle electrode fenestration to removed the stone.6 patients with a single stone were performed with endoscopic sphineterotomy(EST).15 patients are successfully performed with ERCP+EST after cholangitis and general situation turned better.2 patients failed with endoscopic therapy,and they were recovered treated with conventional surgery.Conclusion Minimally invasive intenvention on ASC has characteristics of quick,minimally invasive surgery,high success rate,rapid recover,and fewer complications.

Objective To study the effect of continuing flushing with heparin on the prevention of tube obstruction after vacuum sealing drainage(VSD).Methods Seventy-four patients with large area of skin defects treated by VSD were randomly divided into experiment group and control group. The experiment group were managed by continuous infusion with saline at 250 mL and heparin at 12500 U (50 U/mL)in 24 h after operation,with 20 to 30 drops per minute and two times a day. The control group included 33 cases was injected with 20 mL saline when the drainage tube blocked until the VSD tube was unobstructed. The two groups were compared in terms of drainage time, drainage tube,wound healing time,rate of un-obstruction of tubes and histopathological scores.Results The drainage time and wound healing time of experiment group were significantly shorter than that of control group(P<0.05). The rate of un-obstruction of tubes was higher than the control group and so it was with the score in histopathology.Conclusion Continuous flushing with heparin can effectively reduce the incidence of VSD obstruction and promote wound healing.

Objective To analyze the characteristics and the related risk factors of the contractility of pelvic floor muscles in old women with pelvic floor dysfunction(PFD).Methods A cohort of 125 elderly patients with symptoms of pelvic floor dysfunction,including 60 of postpartum urinary incontinence(SUI)and 65 of pelvic organ prolapse(POP),were recruited.60 women volunteers with normal pelvic floor function were recruited as a control group.A questionnaire survey was conducted to record the general situation of the subjects.The pelvic floor muscle strength was determined by the methods of electromyography and pressure feedback recording.Results The body mass index,the average number of delivery,newborn baby's body weight were significantly higher in the SUI and POP group than in control group,with statistically significant difference(F=5.29,6.27,5.26,P=0.007,0.003,0.008),but without statistically significant difference between SUI and POP groups (P =0.674,0.554,0.578).The rapid contractility value was significant lower in SUI group than in POP and control groups(all P＜0.01),and the sustained contractility value was significant lower in POP group than in SUI and control groups(all P＜0.01).Conclusions The postpartum pelvic floor dysfunction may be correlated with BMI,vaginal delivery rate,the average number of delivery,newborn baby's body weight and a history of increased abdominal pressure.The postpartum pelvic floor dysfunction may be associated with the declined contractility of pelvic floor muscle,among which the POP may be associated with class Ⅰ muscle and the SUI with class Ⅱ muscle.

Objective To investigate the relationship of serum ferritin(SF) and acute coronary syndrome(ACS) .Methods ACS group(n= 110) was divided into unstable angina(UA) subgroup(n= 46) ,non-ST-segment elevation myocardial infarction (NSTEMI) subgroup(n=30) and ST-segment elevation myocardial infarction(STEMI) subgroup(n=34) according to clinical da-ta ,coronary angiography and electrocardiogram examination .The normal coronary patients provided as control group(n=42) .The levels of serum ferritin ,hs-CRP and serum lipids were determined and the differences of SF among the groups were analyzed .The correlation of hs-CRP ,TG ,LDL-C ,HDL-C ,TC and SF were also analyzed .Results Compared with control group ,the level of SF in UA ,NSTEMI ,STEMI were significantly increased(P<0 .05) .Comparison in the levels of SF in NSTEMI ,STEMI group and UA group had significant difference(P<0 .05) .Correlation analysis indicated that the level of SF were positively correlated with hs-CRP ,TG and negatively associated with HDL-C .Conclusion The level of SF significantly increased in patients with ACS and has a positive relationship with hs-CRP and the disorder of lipid metablism .SF helps in assessing the clinical classification and risk strati-fication in patients with ACS .

Objective To investigate the effect of pretreatment with U75302,antagonist of leukotriene B4 receptor 1 (BLT1),on cisplatin induced acute kidney injury in mice and its immunoregulatory mechanism.Methods Healthy C57BL/6 mice were randomized into four subgroups:1.healthy control group;2.cisplatin group;3.U75302 control group;4.cisplatin + U75302 group,n=6.Group 2 and 4 received intraperitoneal injection of cisplatin (20 mg/kg) on day 0,group 3 and 4received intraperitoneal injection of U75302 (5 μg/mouse) on day 0 and day 2.Mice were sacrificed on the 3rd day and blood and kidney were collected.Renal function and histological changes were estimated,the infiltration of immune cells were determined by flow cytometry,the level of peroxidase (MPO) in kidney were determined by colorimetry,relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were detected by Real-time PCR.Results Compared with healthy control group,levels of BUN,Scr were higher in cisplatin group with serious tubular structural damage.There were more neutrophils,macrophages,CD4+ T lymphocytes,CD8+ T lymphocytes in kidneys of cisplatin group,the level of MPO and relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were also higher in cisplatin group.Compared with cisplatin group,lower BUN [(17.75±1.80) mmol/L vs (42.6±6.66) mmol/L,P ＜0.05],Scr were found in cisplatin+ U75302 group with less tubular structural damage.Meanwhile,U75302 reduced infiltration of neutrophils [(146±13)×103/g vs (296±66) ×103/g,P ＜ 0.05],macrophages [(245± 13)× 103/g vs (420±78)× 103/g,P ＜ 0.05] in the kidney.Levels of MPO [(1.756±0.283) U/g vs (3.308±0.577) U/g,P＜0.05] and relative expression of TNF-α,IL-1β,CXCL1,CXCL2 were also lower.Conclusions BLT1 antagonist U75302 protects mice against AKI induced by cisplatin,and the mechanism is associated with reduced infiltration of inflammatory cells in kidney and the inhibition of kidney inflammation.

Objective To establish blood purification system for rats and provide a safe and reliable experimental platform for further research of blood purification. Methods The right carotid artery and the contralateral jugular vein of adult male Sprague?Dawley rats were cannulated to creat vascular access for blood purification, by which continuous arteriovenous hemofiltration blood purification system was established. Blood flow, substitution fluid flow and ultrafiltration rate were regulated by rotary mini?pumps. Blood purification therapy continued for 4 hours on the basis of maintained anesthesia and effective anticoagulation. The safety of continuous arteriovenous hemofiltration blood purification systems was evaluated by comparing the arterial blood gas, electrolyte indexes and blood glucose during the blood purification therapy. Closely monitoring the vital signs of rats, such as blood pressure and heart rate, and observing whether there were any side effects, such as massive haemorrhage, thrombogenesis and gas embolism in the therapeutic process. Results There were no obvious changes of arterial blood gas, electrolyte indexes and blood glucose during the blood purification therapy (P>0.05). The vital signs did not fluctuate acutely before and after the blood purification therapy (P>0.05). The incidence rate of side effects was very low. Conclusions
Continuous arteriovenous hemofiltration blood purification system had no obvious adverse effects on healthy rats. Our blood purification system for rats appears to be safe and reliable.

ObjectiveTo reproduce a clinically relevant two-hit model of sepsis complicated by pneumonia and to explore the correlation between two-hit and immune state.Methods Eighty-one male Sprague-Dawley ( SD ) rats were divided into groups according to the random number table. Forty-five male rats were assigned respectively to sepsis-alone group, pneumonia 4 days and 7 days after sepsis groups, respectively. Survival rate of each group was observed. Another group of 36 male rats were divided into normal control group, sepsis-alone for 1, 4 and 7 days groups, and sepsis complicated by pneumonia for 4 days and 7 days after sepsis groups, each group consisted of 6 rats. Cecal ligation and puncture (CLP) was done in rats, andStreptococcus pneumoniae suspension (bacteria count 1×1010 cfu/mL) was injected via the nose on the 4th day or 7th day after CLP. Rats were sacrificed at corresponding time points, and 1 day after challenge ofStreptococcus pneumoniae on the 4 days or 7 days post CLP for the collection of blood and tissue samples to make bacterial count of the blood, splenocyte count, biochemical indices, cytokines concentration, pathological changes in spleen and apoptotic cells.Results① Compared with the rats of sepsis-alone group, the rats in pneumonia 4 days after CLP group had poor survival rate (4 vs. 11,χ2 = 6.533,P = 0.011), while no difference was found between pneumonia 7 days after CLP group and sepsis-alone group (9 vs. 11,χ2 = 0.600,P = 0.439).② The blood bacterial count and all the biochemical indexes were sharply increased on 1 day post-CLP in the rats of sepsis-alone group, and then they gradually lowered. Compared with the rats of 1 day post-CLP, the proportion of splenocytes were decreased on the 4th day post-CLP [dendritic cells (DC): (0.69±0.09)% vs. (0.87±0.31)%, CD4+T cells: (21.05±2.89)% vs. (24.84±4.59)%, CD8+ T cells: (10.62±1.79)% vs. (13.40±1.31)%, allP< 0.05], but T-regulatory cell (Treg) count was higher on the 4th day after CLP compared with sepsis-alone rats [(3.14±0.74 )% vs. (2.87±1.08)%,P< 0.05]. The biochemical indices, including alanine transaminase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and serum creatinine (SCr) were obviously lowered on 7 days post-CLP compared with 1 day after CLP [ALT (U/L): 35.33±11.52 vs. 81.00±38.40, AST (U/L): 70.33±42.16 vs. 156.00±28.11, BUN (mmol/L): 5.30±2.27 vs. 9.13±4.04, SCr (μmol/L): 55.33±10.67 vs. 96.67±45.79, allP< 0.05]. The serum levels of tumor necrosis factor-α (TNF-α) and interleukins (IL-6, IL-1β) peaked on the 1st day after CLP [TNF-α:(18.03±2.88) ng/L, IL-6: (10.37±4.20) ng/L, IL-1β: (102.44±51.46) ng/L], and high mobility group box-1 (HMGB1) peaked on the 4th day after CLP [(1.76±0.71)μg/L]. The levels of anti-inflammatory cytokines transforming growth factor-β1 (TGF-β1 ) and soluble tumor necrosis factor receptor-Ⅰ (sTNFR-Ⅰ) maintained at high levels [7 days post-CLP: TGF-β1 was (0.90±0.56) ng/L, sTNFR-Ⅰ was (1.56±0.39) ng/L]. The spleen pathology became more marked with the time in the group of sepsis-alone, meanwhile the number of apoptotic spleencytes increased 4 days post-CLP as compared with that of the 1st day post-CLP (cells/HP: 52.99±20.79 vs. 16.05±3.28,P< 0.05).③ Compared with the same period of sepsis-alone group, the rats with pneumonia 4 days post-CLP group showed a higher blood bacterial count (log cfu/mL: 1.78±0.54 vs. 0.25±0.18,P< 0.05), while no difference was found between 7-day of post-CLP pneumonia group and sepsis-alone group (log cfu/mL: 0.57±0.46 vs. 0.13±0.12,P> 0.05). The same trend of changes, with slight reduction in splenocytes and biochemical indices were found between the groups of sepsis followed by pneumonia and sepsis-alone, but no significant difference was found. The level of HMGB1 in the 4-day group of sepsis with complication of pneumonia was further decreased compared with sepsis-alone group (μg/L:1.17±0.74 vs. 1.76±0.71,P< 0.05), and IL-1β in the 7-day group of sepsis complicated pneumonia was further higher than those of sepsis-alone group in the same period (ng/L: 105.73±25.06 vs. 61.04±31.29,P< 0.05), while there were no differences in levels of other cytokines between two-hit group and sepsis-alone group. Apoptosis of spleencytes in the 4-day group of sepsis complicated pneumonia was more marked than that of sepsis-alone group at the same period (cells/HP: 74.48±22.47 vs. 52.99±20.79,P< 0.05), while no difference was found between the 7-day groups of sepsis complicated pneumonia and the sepsis-alone group (cells/HP: 28.70±4.13 vs. 30.43±14.55, P> 0.05).Conclusions The mortality of this two-hit model with complication of pneumonia 4 days after CLP was significantly higher than that of single sepsis model. The ability of bacteria clearance was decreased, and immunocyte apoptosis was exacerbated. These findings may be with the result of the occurrence of immunoparalysis in the mid stage of sepsis. The two-hit model reproduced on 7 days after CLP might suggest reconstruction of host immune function, and maybe associated with the recovery of immune response.

Objective To study the anti-calcification function properties of bovine jugular conduit with valves stabilized by dye-mediated photooxidation.Methods Sixteen bovine jugular conduit with valves were divided into two groups and treated with dye-mediated photooxidation(groupⅠ) and glutaraldehyde(group Ⅱ).The bovine jugular vein was cut into pieces and implanted subcutaneously in the 16 weanling SD rats.After 90 days,all the rats were sacrificed and the retrieved specimens were undergone histological examination by electron microscope and microscope.The calcium content was determined by flame atomic absorption spectrophotometer.Results The walls and valves of bovine jugular vein treated by dye-mediated photooxidation had less calcification than those of the group Ⅱ.Conclusion The dye-mediated photooxidation can effectively preserve the calcification of bovine jugular conduit with valves compared with the way treated by glutaraldehyde.

Aim To research dynamically the changes of endogenous cystathionine- β-synthase/hydrogen sul-fide system in PC12 cells injury induced by rotenone. Methods Rotenone-induced injury in PC12 cells ( characteristic of dopaminergic neurons) was used as a PD cell model. The expression of CBS was evaluated by Western blot. Intracellular CBS activity and H2 S production were detected by Methylene blue spectro-phot-ometric method. The viability of PC12 cells was measured by CCK-8 assay. GSH detection kit was used to detect the intracellular GSH content. Results In the groups of 6 and 12 hours, the expression and activ-ity of CBS were elevated, and the production of H2 S was increased. In the groups of 24 and 48 hours, CBS expression and activity were significantly decreased, and the amount of H2 S was significantly reduced. Ap-plication of 1. 5 μmol·L-1 rotenone for different time (6-48h) could decrease the cell viability and intra-cellular GSH contents in a time-dependent manner. Conclusions The expression and activity of endoge-nous CBS, stimulated by rotenone, are elevated firstly and then decreased. The generation of H2 S, stimulated by rotenone, is increased and then reduced significant-ly, which may be related to PC12 cells against oxida-tive stress damage induced by rotenone.