To study the enrichment regulation of anammox bacteria during the whole start-up process of anammox reaction, two reactors with addition of carries of Spherical Plastic (SP) and Bamboo Charcoal (BC) and one without carrier (CK) were used to start anammox reaction. Then FISH and q-PCR analyses for the growth of all anammox bacteria were conducted during the operational process. The results indicate that the number of anammox bacteria in all reactors increased with time during the whole start-up process, which was consistent with the removal rate of ammonium and nitrite. On day 123 of stable phase, the percent of anammox cells in the sludge of CK, SP and BC accounted for 23.3%, 32.6% and 43.7%, respectively. The number of anammox bacteria 16S rRNA gene copies was (25.64 +/- 2.76) x 10(7), (47.12 +/- 2.76) x 10(7) and (577.99 +/- 27.25) x 10(7) copies g(-1) VSS in the sludge of CK, SP and BC, respectively. Carrier addition could dramatically increase enrichment of anammox bacteria. BC addition significantly increased the anammox bacteria number in the UASB reactor which resulted in the acceleration of the anammox start-up process. In addition, the max specific growth rate and the minimum doubling time were 0.064 d(-1) and 10.8 d in BC reactor. The max specific growth rate of anammox bacteria in BC reactor was 1.78 times and 1.88 times greater than that in CK and SP reactor, respectively. Therefore, the FISH and q-PCR analyses were suitable for determining the enrichment regulation of anammox bacteria during the start-up time, while a bit of differences in results existed between the two analytical methods due to the difference in analysis targets.
Ammonia ; metabolism ; Bacteria ; growth & development ; metabolism ; Bioreactors ; Industrial Microbiology ; Nitrites ; metabolism ; Oxidation-Reduction ; Sewage ; microbiology

In order to simultaneously remove carbon and nitrogen from organic-rich wastewater, we used an up-flow anaerobic sludge bed/blanket (UASB) reactor that was started up with anammox with high concentration of carbon and nitrogen by gradually raising the organic loading of influent. We optimized the removal of nitrogen and carbon when the chemical oxygen demand (COD) concentration varied from 172 to 620 mg/L. During the entire experiment, the ammonium and total nitrogen removal efficiency was higher than 85%, while the average COD removal efficiency was 56.6%. The high concentration of organic matter did not restrain the activity of anammox bacteria. Based on polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and tapping sequencing analyses, the Planctomycete, Proteobacteria, Chloroflexi, Chlorobi bacteria are detected in the UASB reactor, which indicated complex removal pathway of carbon and nitrogen coexisted in the reactor. However, a part of Planctomycete which referred to anammox bacteria could tolerate a high content of organic carbon, and it provided help for high performance of nitrogen removal in UASB reactor.
Ammonia ; chemistry ; Biological Oxygen Demand Analysis ; Bioreactors ; Carbon ; chemistry ; Nitrogen ; chemistry ; Sewage ; Waste Disposal, Fluid ; methods ; Waste Water ; chemistry

OBJECTIVE:To establish the quality standard of Belamcandin standard substance.METHODS:Different indices of standard Belamcandin were examined with quantitative and quantitative analyses according to the corresponding measures stipulated in the Appendix of pharmacopoeia of people's republic of China(part Ⅱ)published in 2005.RESULTS:The specific rotatory power of 3 batches of standard Belamcandin was —28?～—30?([?]25 D,c0.11,Pyridine);the E1% 1 cmwas 790～830;the melting point was 252～254 ℃ and the ash content was less than 0.5%.The content of Belamcandin in the products was above 99.5% as revealed by HPLC.CONCLUSION:All of the indices were shown to meet the standards for standard substances and the established standard can be used for the quality control of standard Belamcandin.

Objective To explore the impact of nutritional risk on short-term clinical outcomes after laparoscope-assisted radical gastrectomy for gastric cancer.Methods The retrospective case-control study was conducted.The clinical data of 150 patients who underwent laparoscopic gastrectomy at the First Affiliated Hospital of Wenzhou Medical University between June 2014 and April 2016 were collected.According to nutritional risk screening 2002 (NRS 2002),42 and 108 patients were respectively divided into the nutritional risk group (NRS 2002 score ≥3) and non-nutritional risk group (NRS 2002 score ＜3).Laparoscope-assisted radical subtotal gastrectomy or total gastrectomy was performed based on tumor location.Observation indicators:(1) postoperative short-term clinical outcomes:postoperative complications,duration of postoperative hospital stay,hospital expenses,unplanned readmission within 30 days after discharging.Postoperative complications meant total complications within 30 days postoperatively,grade Ⅰ-Ⅴ of Clavien-Dindo grade was complication classification.Grade Ⅱ and above of Clavien-Dindo grade were analyzed in this research.(2) Risk factors analysis affecting occurrence of postoperative complications of patients.Measurement data with normal distribution were represented as x±s and analyzed using the independent-sample t test.Measurement data with skewed distribution were described as M (Qn) and analyzed using the Mann-Whitney U test.Categorical variables were described as number and percentage and analyzed by the chisquare test.Ranked data were analyzed by the Mann-Whitney U test.Univariate analysis was done by the chi-square test.P＜0.1 of univariate analysis was used to multivariate analysis.COX regression model in multivariate analysis was built using progressive condition method.Results (1) Postoperative short-term clinical outcomes:number of patients with total complications,number of patients with severe complications,duration of postoperative hospital stay,hospital expenses and number of patients with unplanned readmission within 30 days after discharging were 9,2,11 days (9 days,16 days),57 825 yuan (51 894 yuan,66 908 yuan),2 in the nutritional risk group and 16,3,11 days (9 days,13 days),55 067 yuan (49 395 yuan,62 423 yuan),8 in the non-nutritional risk group,respectively,with no statistically significant difference between the 2 groups (X2=0.952,0.010,Z=-1.133,-1.691,X2 =0.048,P＞0.05).Results of univariate analysis showed that age was a risk factor affecting incidence of complications after laparoscope-assisted radical gastrectomy for gastric cancer (X2 =4.468,P＜ 0.05).Results of multivariate analysis showed that preoperative hypoproteinemia was an independent risk factor affecting incidence of complications after laparoscope-assisted radical gastrectomy for gastric cancer (OR =2.797,95％ confidence interval:1.053-7.435,P＜0.05).Conclusion There is little poor impact of nutritional risk on short-term outcomes after laparoscope-assisted radical gastrectomy for gastric cancer,preoperative hypoproteinemia is an independent risk factor affecting occurrence of grade Ⅱ and above of postoperative complications.

To ensure the quality and safety of Panax notoginseng, a method for the simultaneous determination of 10 mycotoxins in Panax notoginseng was developed using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The sample was extracted with acetonitrile and purified by HLB multifunction cleanup column. The separation was performed on a Phenomenex Kinetex XB-C18 column by gradient elution using methanol and 5 mmol·L(-1) ammonium acetate as mobile phase. The targeted compounds were detected in MRM mode by mass spectrometry with electrospray ionization (ESI) source operated in both positive and negative ionization modes. The linear relationships of the 10 mycotoxins were good in their respective linear ranges. The correlation coefficients (r) ranged from 0.9981 to 1.0000. The LOQs of the 10 mycotoxins were between 0.15 and 8.6 μg·kg(-1). The average recoveries ranged from 73.8% to 107.0% with relative standard deviations (RSDs) of 0.10%-10.9%. The results demonstrated that the proposed method was sensitive and accurate, and suitable for the mycotoxins quantification in Panax notoginseng.

Objective To improve the 2,3,5-triphenyl tetrazolium chloride (TTC) solution method for measuring the viability of Cistanche deserticola seeds and investigate the change in viability during storage at 5 ℃. Methods The effect of the testa,TTC concentration,sodium hypochlorite concentration (NaClO),and staining time were studied,and seed viability during storage at 5 ℃ was measured with the improved method. Results Seeds were kept for 48 h in 0.5% TTC solution at 40 ℃,and then for 2 h in 0.2% NaClO solution;Seed viability was measured under a stereomicroscope. Storing seeds of C. deserticola for 1 to 2 years at 5 ℃ had no significant effects on their viability. However,the percentage of seeds with high viability was increased with the extension of the storage time at 5 ℃. Conclusion A convenient and rapid method for measuring the viability of C. deserticola seeds is developed. Storing C. deserticola seeds at 5 ℃ could improve their viability

Objective:By comparing preoperative high resolution magnetic resonance imaging (MRI) examination with postoperative pathologic results, to investigate the effects of MRI examination evaluation on the anatomical level and clinical outcome.Methods:We conducted a retrospective study on 72 patients who underwent resection of rectal cancer at the Second Affiliated Hospital of Wenzhou Medical University between Apr 2017 and Nov 2018, including 35 patients undergoing laparoscopic resection and 37 patients doing open resection. All cases received high resolution MRI examination before operation. The diagnostic accuracy of MRI, operation safety, and the short-term outcomes were analyzed.Results:There were no postoperative tumor recurrence. The accuracy rate of preoperative MRI evaluation of T stage was 85%, and positive N+ was 74%. There were no difference in postoperative complications between the open resection group and laparoscopic resection group (29% vs. 22%, χ 2=0.463, P=0.496). The proximal and distal margin was negative, postoperative circumferential resection margin and preoperative mesorectal fascia was consistent, the distance between the lower margin of the tumor and the anal right angle measured by MRI were consistent with the distance between the tumor from the dentate line. Conclusion:High resolution MRI with a good tissue resolution, has a high preoperative diagnosis accuracy for T and N staging of the low rectal cancer, with decisive role in the evaluation on the anatomical level, improving the quality and safy of surgery.

OBJECTIVETo explore the central mechanism of postoperative fatigue syndrome by detecting the expression of NMDA receptor and tryptophan metabolism.

METHODSAfter being numbered according to the weight, ninety-six male SD rats were randomly divided into control group (bowel loop was flipped after laparotomy and received intraperitoneal injection of saline at a dose of 1 ml/kg), POFS model(70% of the length of small intestine was resected and received intraperitoneal injection of saline at a dose of 1 ml/kg), and NMDA antagonist groups(70% of the length of small intestine was resected and received intraperitoneal injection of MK801 at a dose of 1 ml/kg). Each group was divided into subgroups by postoperative 1, 3, 5 and 7 d, with 8 rats in each subgroup. The hippocampus was removed at each time point after open field test (OFT) to detect the mRNA expression levels of NMDA receptor 1 and kynurenine aminotransferase III((KATIII() by real-time PCR. Protein level of NMDA receptor 1 was detected by Western blot. High performance liquid chromatography (HPLC) was used to measure the concentrations of tryptophan (TRP), kynurenine (KYN) and kynurenic acid(KYNA). Ultra-structural changes of hippocampal neurons were observed by transmission electron microscopy(TEM).

RESULTSAs compared to control group, exercise score decreased(P<0.05), rest time and central panel residence time prolonged, periphery/central panel ratio increased (all P<0.05), mRNA and protein expressions of NMDA receptor 1 increased (P<0.05), mRNA expression of KAT III( decreased (P<0.05), KYN/TRP ratio and KYN/KYNA ratio decreased (all P<0.05) in POFS group on postoperative day 1 and 3. As compared to POFS group, central panel residence time and periphery/central panel ratio decreased on postoperative day 1, and mRNA and protein expressions of NMDA receptor 1 decreased on postoperative day 1 and 3 (all P<0.05) in antagonist group. TEM revealed that degenerated neuron was found in the hippocampus of POFS rats, while such damage was improved in antagonist group.

CONCLUSIONThe increased expression level of NMDA receptor may play an important role in POFS. NMDA receptor antagonist MK801 may improve the POFS.

Animals ; Fatigue ; Hippocampus ; Humans ; Injections, Intraperitoneal ; Male ; Postoperative Period ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; Signal Transduction ; Transaminases

Objective: To investigate the effect of visceral obesity on the short-term outcomes after radical operation for mid-low rectal cancers. Methods: We conducted a prospective study on patients who underwent selective rectal cancer resection at The Second Affili-ated Hospital of Wenzhou Medical University between April 2017 and October 2018. The cutoff visceral fat area (VFA) for visceral obe-sity was≥134.6 cm2 for men and≥91.1 cm2 for women. Results: A total of 127 patients were included in the study, of whom 64 were diagnosed as having visceral obesity and 63 as having non-visceral obesity. The patients with visceral obesity had a higher body mass index (BMI) (P<0.001) than those without visceral obesity. The proportions of female patients and those who had a laparoscopy-assist-ed surgery were higher in the visceral obesity group than in the non-visceral obesity group. We found no significant differences in age, albumin level, hemoglobin count, American Society of Anesthesiologists (ASA) stage, Nutritional Risk Screening (NRS) 2002 score, Charlson comorbidity index, tumor location, TNM stage, lymphatic invasion, and laparoscopy-assisted surgery between the two groups. The postoperative complication rate was significantly higher in the visceral obesity group than in the non-visceral obesity group (35.9% vs . 19% , P=0.033). A multivariate Logistic regression analysis revealed that visceral obesity [odds ratio (OR)=2.732, P=0.019] and NRS 2002 scores of≥3 (OR=2.574, P=0.042) were independent risk factors for postoperative complications. Conclusions:Visceral obesity was an independent risk factor for complications after surgery for mid-low rectal cancers.

OBJECTIVE：To establish the method for content dete rmination of polyphyllin Ⅱ，Ⅵ，Ⅶ in Ypsilandra thibetica ， and to compare the differences of 3 saponins in different parts of Y. thibetica from different producing areas. METHODS ：HPLC method was adopted to determine the contents of polyphyllin Ⅱ，Ⅵ，Ⅶ in whole grass part and underground part of Y. thibetica from 10 producing areas. The determination was performed on Kromasil C 18column with mobile phase consisted of acetonirile-water （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was 35 ℃，and detection wavelength was set at 203 nm；sample size was 10 μL. With the contents of 3 saponins as the index ，20 batches of Y. thibetica were analyzed by cluster analysis and PLS-DA analysis ；the aggregation of samples was analyzed and determined the primary difference components. RESULTS：The linear range of polyphyllin Ⅱ，polyphyllin Ⅵ and polyphyllin Ⅶ were 0.051-2.04，0.007-0.28，0.168-6.72 μg， respectively（r≥0.999 5）；the detection limits were 1.92，1.75，1.87 ng，respectively；and the quantitative limits were 6.40，5.87， 6.23 ng，respectively；RSD of precision ，reproducibility and stability tests （24 h）were all lower than 2％（n＝6）；the average recovery rates were 99.29％，101.38％ and 99.64％，with RSDs of 1.17％，2.64％，0.75％（n＝6），respectively. The content of polyphyllin Ⅱ was 0.615-1.875 mg/g，that of polyphyllin Ⅵ was 0-0.095 mg/g，and that of polyphyllin Ⅶ was 3.158-12.354 mg/g. Cluster analysis showed that 20 batches of samples were clustered into two groups ，batch S 9-S12 were clustered in to one group，and the other 16 batches of samples were clustered into another group. PLS-DA analysis showed that 20 batches of samples were divided into 3 areas，batch S 1，S2，S8，S14，S16，S20 were included in area Ⅰ；batch S 9-S12 included in area Ⅱ；and the rest of the samples included in area Ⅲ. The quality of Y. thibetica from different habitats was different ，and there was no difference in the saponin composition between the whole grass and the underground part. Weight ranking found that mail：cscdtcm@126.com polyphyllin Ⅶ was the main difference component in Y. thibetica，and the content of polyphyllin Ⅶ in Y. thibetica from Pengzhou city and Dayi county was the highest. CONCLUSIONS ：The established method is simple ，convenient and sensitive. It can be used for the content determination of 3 saponins in Y. thibetica . The content of active components is higher and the quality is better in Y. thibetica from Pengzhou city and Dayi county.