Abstract A method based on dispersive solid phase extraction-high performance liquid chromatography-tandem mass spectrometry for determination of 19 kinds of nonprotein nitrogen compounds including melamine, cyromazine, amidinourea, aminotriazine, 3-aminotriazole, 4-aminotriazole, allantoin, cyanuric acid, dicyandiamide, thiourea, semicarbazide, L-leucine, L-isoleucine, L-arginine, L-hydroxyproline, L-theanine, ammeline, ammelide and guanidine in powdered formulas was presented. The nonprotein nitrogen compounds were degreased by chloroform and extracted by acetonitrile, with MgSO4 to remove water and C18 to clean up. The samples were separated on Merck ZIC HILIC column (150 mm í2. 1 mm, 5 μm, 20 nm) with gradient elution. The electrospray ionization was operated in the positive mode and negative mode, and monitored by the multiple reaction monitoring ( MRM) mode. Allation was quantified by external standard method and the other 18 kinds of nonprotein nitrogen compounds were quantified by internal standard method. All of the correlation coefficients (r) were higher than 0. 99. The limits of quantitation (LOQ) were 0. 05-5. 0 mg/kg, the average recoveries were between 82 . 2% and 115 . 0%, and the relative standard deviations were less than 20%.

Objective:To explore the effect of ulinastatin on prevention of acute respiratory distress syn-drome (ARDS).Methods:A prospective multicentral cohort study was conducted.The patients from three intensive care units (ICUs)of grade A tertiary hospitals in Beijing and a ICU of grade A tertiary hospitals in Cangzhou from January 2012 to December 2014,included 77 ARDS at-risk patients with uli-nastatin treatment and 108 ARDS at-risk patients without ulinastatin treatment (control)were eligible. Both groups received normal treatment;additionally,the intervention group received 600 000 units of uli-nastatin via intravenous infusion for 5 days.The control group received the same amount of saline via in-travenous infusion for 5 days.Venous blood human neutrophil elastase (HNE)and peptidase inhibitor 3 (PI3)levels were measured on days 1,3,and 7,respectively.Other outcomes included acute physiolo-gy and chronic health evaluation scoring Ⅱ (APACHE Ⅱ),body temperature,respiratory rate,heart rate,mean arterial pressure,white blood cell counts,PaO2 /FiO2 ,ARDS incident,mechanical ventila-tion time,ICU treatment and hospitalization duration,28 days mortality.Results:The PI3 levels showed no statistical difference on day 1,but significant differences on day 3 and day 7 between the two groups (P <0.01).HNE /PI3 ratio showed no statistical difference on day 1,but significant differences on day 3 and day 7 (P <0.05).PaO2 /FiO2 was significantly higher in ulinastatin group on day 3 and day 7 (P <0.05).The incident rate for ulinastatin group was 15.58%,lower than that for the control group (33.33%),and the difference was statistically significant (P <0.05).The mechanical ventilation time and ICU treatment time in ulinastatin group was shorter than that in the control group,and the difference was statistically significant (P <0.05).There were no significant effects in other factors.Conclusion:Increased dose of ulinastatin can recover the balance of HNE and its antagonist,lower the HNE’s damage to lungs,and further reduce the ARDS incident rate.

Objective To observe the distribution and possible mechanism of P2X7R in periaq-ueductal gray matter (PAG)in a rat model with chronic neuropathic pain in vivo.Methods The in-trathecal catheterization and sciatic nerve injury (SNI)were performed.All animals were randomly assigned into 3 groups with 26 rats in each,which was group Sham,control group (group C)and brilliant blue G (BBG)group (group BBG),respectively.Normal saline or BBG 10 μl were intrathe-cally injected after SNI and repeated for seven days.Paw-withdrawal mechanical thresholds (PWT) were measured on day 0,day 7,day 14,and day 21 after SNI.The rats were sacrificed and PAG tis-sues were collected on day 14 and day 21,separately.The distributions of P2X7R were observed by immunofluorescence.The protein contents of P2X7R and GFAP were assessed by Western blot assays.Results The P2X7R was expressed in PAG in rats.The PWTs of the control group showed a significant decrease during the 21-day period compared with the sham group.The P2X7R signals were predominantly expressed in astrocytes in PAG after SNI.Both P2X7R and GFAP expression remark-ably increased.Administration of BBG increased the PWTs,and inhibited the P2X7R and GFAP ex-pressions compared with those atthe same point of time of the control group.Conclusion These results indicated that P2X7R in PAG might participate in nociception modulation in the midbrain in chronic neuropathic pain.

Objective To investigate the incidence and risk factors of surgical site infection ( SSI ) in patients with colorectal cancer .Methods Clinical data of patients with colorectal cancer undergoing surgical treatment in Jiaxing First Municipal People’ s Hospital from October 2011 to December 2014 were retrospectively reviewed.The gender, age, underlying diseases, smoking history, preventive medication, abdominal surgery history , type of surgery , preoperative levels of hemoglobin and albumin , use of laparoscopy, use of stapler, combined organ resection, TNM staging, American Society of Anesthesiologists ( ASA) score was documented .Multivariate logistic regression analysis was performed to identify the risk factors of SSI .Results A total of 773 patients were enrolled in the study , and SSI was observed in 144 cases (18.63%).Multivariate logistic regression analysis showed that use of laparoscopy ( OR =0.35, 95%CI:0.15-0.79,P <0.05), use of stapler (OR =0.59, 95% CI: 0.39-0.88,P <0.05) were protective factors for SSI, while diabetes (OR=2.11, 95% CI: 1.25-3.58,P<0.01), liver cirrhosis (OR=2.12,95%CI:1.18-3.79,P<0.05), ASA score (3-4 points) (OR=2.01,95%CI:1.20-3.58, P<0.01), combined organ resection (OR=2.17,95% CI:1.20-3.92,P<0.05), and anastomotic leak (OR=6.85, 95%CI:3.01-15.63,P<0.01) were risk factors for SSI.Conclusions The incidence of SSI is high in patients with colorectal cancer undergoing surgery .Use of laparoscopy and stapler may reduce the incidence of SSI .

Objective To evaluate the incidence and significance of perineural invasion (PNI) in cervical cancer. Methods Retrospective chart review of patients with cervical cancer (stages Ⅰa2-Ⅱb) who underwent radical hysterectomy and pelvic lymphadenectomy from 2007 to 2012. To evaluate the incidence and significance of PNI in cervical and uterine tissues by microscopic examination. Results A total of 238 patients were included, 9.2% (22/238) patients with PNI in the cervical stroma. Patients with PNI were more likely to have adverse histopathologic features, including lymphoma vascular space invasion, parametrical invasion, depth of invasion, tomor size and lymph nodes metastases (all P<0.05). PNI were independent of age, international federation of gynecology and obstetrics (FIGO) stage, histopathology type and grade, and positive vaginal margin (all P>0.05). Patients with PNI had shorter disease-free and overall survival (P=0.002 and P=0.008, respectively). On multivariate analysis, risk factors for recurrence and death included parametrical invasion and depth of invasion (P<0.05). Similarly, risk factors for recurrence included lymph nodes metastases (P=0.024). However, PNI was not identified as an independent risk factor for either recurrence or death (P>0.05). Conclusions PNI exists in early cervical cancer. PNI is associated with tumor size, depth of invasion, parametrical invasion, lymphoma vascular space invasion and lymph nodes metastases. PNI represente a decreasing disease-free and overall survival in patients with early-stage cervical cancer, and is independently associated with multiple high-risk factors, which be informed management decisions regarding adjuvant therapy.

OBJECTIVE: To establish a hepatocyte immunotoxicity model for screening of liver protective medications. METHODS: Cytotoxicity was induced by coincubating BCG-pretreated rat hepatocytes in vivo and with 10 mg/L LPS in vitro. Biphenyldimethylesterate (DDB), malotilate(MLT), silybin(SB) and glycyrrhizin (GRZ) were coincubated along with LPS to prevent the hepatocyte injury and verify the applicability and reliability of the model. AST, LDH and nitric oxide (NO) were measured in both the serum and supernatant. The liver and spleen index were calculated and the liver histopathologic changes were examined microscopically. RESULTS: Supernatant AST, LDH and NO in the BCG combined LPS group were increased in comparison with the control group (P<0.01). This increase was attenuated by the addition of DDB, MLT, SB and GRZ (P<0.05). The serum AST, NO and liver and spleen index were also increased significantly compared with the control group (P<0.01). Microscopic exam revealed serious histopathologic changes in the BCG combined LPS group. Hepatoxicity with associated liver enzyme elevation but histopathologic changes were attenuated by DDB, MLT, SB and GRZ. CONCLUSION: BCG combined LPS-induced hepatocyte immunotoxicity in an in vitro rat model may be a useful technique to assess the effectiveness of liver protective medications.

OBJECTIVETo investigate the expression of long non-coding RNA-HOTAIR in prostate cancer cells and its effects on the growth and metastasis of the cells.

METHODSUsing quantitative reverse-transcription PCR (qRT-PCR), we determined the relative expression of HOTAIR in the normal human prostate epithelial cell line RWPE-I and prostate cancer cell lines PC-3 and DU145. We detected the effects of HOTAIR on the cell cycle and invasiveness of prostate cancer cells by RNA interference, flow cytometry, and Transwell mitration assay.

RESULTSThe expressions of HOTAIR in the PC3 and DU145 cells were increased 3.2 and 5.7 times, respectively, as compared with that in the normal RWPE-1 cells. After si-HOTAIR interference, the prostate cancer cells were arrested in the G2 phase and downregulated in the G1 phase. The invasive ability of the prostate cancer cells was evidently inhibited, with the inhibition rates of 32% and 44% of the PC3 cells and 43% and 34% of the DU145 cells for si-HOTAIR1 and si-HOTAIR2, respectively.

CONCLUSIONIncRNA HOTAIR is highly expressed in prostate cancer, which is associated with the growth and invasiveness of prostate cancer cells. HOTAIR is potentially a novel marker for the diagnosis and prognosis of prostate cancer.

Cell Cycle ; Cell Cycle Checkpoints ; Cell Division ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; G1 Phase ; G2 Phase ; Humans ; Male ; Neoplasm Invasiveness ; Prognosis ; Prostatic Neoplasms ; metabolism ; pathology ; RNA Interference ; RNA, Long Noncoding ; metabolism ; RNA, Untranslated ; metabolism

Objective To understand the epidemiological distribution and epidemic situation of brucellosis cases in Zhejiang Province in 2003 - 2012. Methods Questionnaires of confirmed brucellosis cases, annual reports of prevention and control work of brucellosis in the cities and monitoring points were collected and analyzed in 2003 - 2012. Population distribution, regional distribution, infection sources and routes of infection, aetiology and clinical symptoms and signs of the brucellosis cases were analyzed descriptively. Results A total of 323 brucellosis cases were reported in 2003 - 2012, the average annual incidence rate was 0.070/one hundred thousand, the incidence rate in 2012(0.190/one hundred thousand) was high. There were 272 people of the 323 brucellosis cases were from occupational populations, accounting for 84.21%(272/323); people worked in buying, slaughtering and trafficking of livestock products were majority of the cases, accounting for 65.02%(210/323); and unoccupational population was accounting for 15.79%(51/323). Regional distribution of brucellosis was gradually spreading; the average incidence rate of Shaoxing City was the highest ( 0 . 226/one hundred thousand ) . Three hundred patients were infected by sheep, accounting for 92.88%(300/323), and only 7.12%(23/323) of the patients were infected by cattle. Two hundred and eighty-seven acute phase patients of brucellosis were checked by blood culture, and thirty-nine Brucella melitensis strains and two Brucella abortus strains were detected. The chronicity survey of 120 confirmed cases of brucellosis were conducted, chronicity rate was 3.33%(4/120). Conclusions Trends in the epidemic situation of brucellosis in Zhejiang Province has continued to spread. The main source of infection is sheep from the North. Occupational populations are major populations at risk. The key of prevention and control is to strengthen the active monitoring of occupation personnel , health education and behavior intervention.

Objective To observe the effect of Valpar 4 system on hand injury. Methods 40 patients with hand injury were randomly divided into control group and observation group with 20 patients in each group. Routine treatment was applied in both groups while Valpar 4 system was added in the observation group. Then total active motion (TAM) of finger joints, Disability of Arm Shoulder and Hand (DASH), and Upper Extremity Function Test (UEFT) were carried out before, 2 weeks and 4 weeks after treatment. Results The TAM improved significantly in the control group 2 weeks after treatment (P<0.001). The TAM, DASH and UEFT improved significantly in the observation group (P<0.001), and the DASH and UEFT were better in the observation group than in the control group (P<0.001). 4 weeks after treatment, the TAM and UEFT improved significantly (P<0.001) in the control group, the TAM, DASH, UEFT significantly improved in the observation group (P<0.001), and were better in the observation group than in the control group (P<0.05). Conclusion Valpar 4 system can effectively improve the TAM of fingers, function of hands and upper limb, especially in terms of activities of daily living of upper limbs.