To accurately analyze metabolites in industry-important photosynthetic microbes, LC-MS based metabolomics protocol needs to be optimized specifically for individual species. In this study, an LC-MS based metabolomics method was optimized for cyanobacterium Synechocystis sp. PCC 6803. With the optimized extraction, liquid chromatographic and mass spectral parameters, the method was capable of detecting 24 important metabolites related to central carbohydrate and energy metabolism in Synechocystis sp. PCC 6803. The study laid an important foundation for the metabolomics analysis of cyanobacteria.
Chromatography, Liquid ; Mass Spectrometry ; Metabolome ; Metabolomics ; Photosynthesis ; Synechocystis ; metabolism

Objective: To analyze the features of Norovirus outbreaks in schools and kindergartens and the associated factors in Chengdu, 2017, and to provide the scientific basis for Norovious outbreaks prevention and control. Methods: A total of 75 schools and kindergartens in Chengdu of 2017 participated in the study about Norovirus outbreaks. ANOVA and binary Logistic regression models were used to analyze the influencing factors and duration of Norovirus outbreaks. Results: Overall 710 cases were included. There was an average of 9.47 cases and 27.52 hours for each outbreak,decreasing by 1.06 cases and 10.56 hours compared with those of 2016. Most outbreaks happened in kindergartens and in the firstlevel economy regions from January to March, with GII type as the main outbreak. Compared with the first case of vomiting at home, vomiting in public area (OR=11.76, 95%CI=1.63-84.69) was much more serious, and compared with the active report of school/ community, being informed of the outbreak passively (OR=4.09，95%CI=1.04-16.03) was positively associated with outbreak severity. Conclusion To prevent Norovirus infection, specific development and training of dealing with vomiting and feces should be introduced, and measures to increase the ability to surveil and report Norovirus outbreaks should be enhanced.

Objective: To explore the associations of Hand-Foot-Mouth disease (HFM) among children of different age groups in kindergartens, and to provide scientific evidence for HFM prevention and control. Methods: Cluster sampling was used to select 9 912 children from 40 kindergartens. The major caregivers of children were invited to participate into the study and fulfill a self-administered questionnaire that consisted of general background, hand washing and the history of HFM among children in the recent 1 year. Chi-square and binary logistic regression were applied,to analyze the influencing factors of HFM among children of different age groups. Results: In the 2 year-old-group, children who were male (OR=1.76, 95%CI=1.24-2.50), had the major caregiver with high school educational background or below (OR=1.54, 95%CI=1.06-2.24), had family income more than 100 000 yuan or more per year(OR=1.49, 95%CI=1.01-2.20), children whose major caregiver seldom wash hands immediately after coming home(OR=2.10, 95%CI=1.05-4.19)，and 3-5 times per week(OR=2.07, 95%CI=1.26-3.41) were more likely to have HFM. In the 3 year-old-group, the more time children spent in the outdoors, the less likely they got HFM(3-5 times per week: OR=0.58, 95%CI=0.43-0.81; ≥6 times per week: OR=0.45, 95%CI=0.29-0.70). Children whose major caregiver did not use soap were more likely to have HFM(OR=1.67, 95%CI=1.11-2.49). In the 4 year-old-group, children who occasionally took toys when going out were less likely to get HFM (vs always, OR=0.57, 95%CI=0.38-0.88). No significant factors were found in the 5 year-old-group. Conclusion The behaviors and activities of major caregivers and children could influence the HFM incidence.To prevent HFM, it was advised that the major caregivers should wash hands immediately after coming home, and develop the habit of using soap for handwashing together with children at the earlier age, and encourage children to take more outside-door activities according to the age characteristics. Attention should be paid to avoid contaminating takeout snacks, and to clean the takeout toys timely.

Objective: To analyze the relationship between the toilet design and sanitary condition of primary and secondary schools in Chengdu and the epidemic situation of Norovirus infection, so as to provide scientific references for school toilet design and Norovirus prevention and control. Methods: A total of 78 cases of norovirus epidemics from 2019 to 2020 were included in this study. These epidemics happened in 78 schools and the causes were indicated as human-to-human transmission. The Chi square and Mann-Whitney U test were used to compare categorical and continuous data respectively. Unconditional binary Logistic regression was used to examine the multivariate associations. Results: Most epidemics happened from October in 2019 to March in 2020(79.49%, 62 cases), in primary schools (71.79%, 56 cases) and in the center area (52.56%, 41 cases). The median of the case number in each epidemic was 10. Overall, 56 schools (71.79%) were equipped with toilets which were flushed independently, and 22 schools (28.21%) were equipped with toilets which were flushed uniformly. There were 27 schools (34.62%) that did not have enough water taps. Logistic regression analysis found that those with toilets flushed uniformly had a stronger epidemic, compared to the schools with toilets flushed independently( OR=5.53, 95%CI=1.63-18.76, P <0.05). Conclusion In order to prevent intestinal infectious diseases , it is suggested that schools should design or reconstruct independent flushing toilets with adequate faucets.

Objective To compare the short-term clinical efficacies of Da Vinci robotic surgical systemassisted and laparoscopy-assisted radical gastrectomy for locally advanced gastric cancer (GC).Methods The retrospective cohort study was conducted.The clinicopathological data of 162 patients who underwent minimally invasive radical gastrectomy for locally advanced GC in the First Affiliated Hospital of Army Medical University between September 2016 and September 2017 were collected.Of 162 patients,65 undergoing Da Vinci robotic surgical system-assisted radical gastrectomy were allocated into the robotic group and 97 undergoing laparoscopyassisted radical gastrectomy were allocated into the laparoscopic group.According to Japanese gastric cancer treatment guidelines,patients with upper GC and with middle or lower GC underwent respectively total gastrectomy + D2 lymph node dissection and distal subtotal gastrectomy + D2 lymph node dissection,and then Billroth Ⅱ or Roux-en-Y digestive tract reconstruction.Observation indicators:(1) surgical and postoperative situations;(2) detection of lymph node;(3) follow-up and survival situations.Measurement data with normal distribution were represented as x±s,and comparisons between groups were analyzed using the t test.Comparisons of count data were done using the chi-square test.Ordinal data were analyzed by the nonparametric test.Results (1) Surgical and postoperative situations:all 162 patients underwent successful surgery,without conversion to laparoscopic or open surgery,and pathological resection margins were confirmed as R0.Volume of intraoperative blood loss,levels of amylase in peritoneal drainage fluid at day 1,2 and 3 postoperatively,levels of serum amylase fluid at day 1,2 and 3 postoperatively were respectively (123±39) mL,(557± 181) U/L,(357± 127) U/L,(183±86) U/L,(181±47)U/L,(123±29)U/L,(85±22)U/L in the robotic group and (142±40)mL,(793±284)U/L,(497±199)U/L,(279±157) U/L,(218±45) U/L,(162±37) U/L,(120±31) U/L in the laparoscopic group,with statistically significant differences between groups (t =-3.015,-2.817,-2.364,-2.132,-2.372,-3.338,-3.720,P＜0.05).Cases with distal subtotal gastrectomy + D2 lymph node dissection and with total gastrectomy + D2 lymph node dissection,cases with Billroth Ⅱ and Roux-en-Y of digestive tract reconstruction,time of distal subtotal gastrectomy + D2 lymph node dissection,time of total gastrectomy + D2 lymph node dissection,cases with anastomotic leakage,pulmonary infection,wound infection or liquefaction and delayed gastric emptying,cases in grading Ⅱ,Ⅲ,Ⅳ and Ⅴ of postoperative complications,time of postoperative drainage-tube removal and duration of postoperative hospital stay were respectively 47,18,40,25,(222±37) minutes,(274±43) minutes,1,1,1,1,2,1,0,0,(6.5-± 1.5) days,(10.0±4.0) days in the robotic group and 74,23,69,28,(213±40) minutes,(262±39)minutes,2,4,1,0,4,1,0,1,(6.9±1.7)days,(10.0±5.0)days in the laparoscopic group,with no statistically significant difference between groups (x2=0.326,1.628,t =1.272,0.960,x2=2.501,Z=-1.342,t=-1.142,-0.115,P＞0.05).One and 1 patients in the robotic and laparoscopic groups who were complicated with esophagus-jejunum anastomotic leakage after total gastrectomy + Roux-en-Y anastomosis were cured by nutrition support therapy using feeding tube placement under gastroscopy,and 1 patient in the laparoscopic group who were complicated with gastrojejunal anastomosis leakage after distal subtotal gastrectomy +Billroth Ⅱ anastomosis received the second surgical exploration and jejunal feeding tube placement.Patients with pulmonary infection,wound infection or liquefaction and delayed gastric emptying were cured by conservative treatment.Levels of amylase in peritoneal drainage fluid and serum amylase fluid at day 1,2 and 3 postoperatively were not higher than 3 times of upper limit of normal,without treatment interventions.(2) Detection of lymph node:overall number of lymph nodes detected in the robotic and laparoscopic groups were respectively 36.82±13.41 and 35.21 ± 11.52,with no statistically significant difference between groups (t =0.786,P＞ 0.05).Results of further analysis showed that numbers of lymph node dissected in the 2nd station and upper region of pancreas in patients undergoing distal subtotal gastrectomy + D2 lymph node dissection were respectively 6.04±3.98,13.51±6.53 in the robotic group and 4.45±3.12,11.40±5.30 in the laparoscopic group,with statistically significant differences between groups (t=2.461,1.986,P＜0.05).Numbers of lymph node dissected in No 7 and 8 groups and upper region of pancreas in patients undergoing total gastrectomy + D2 lymph node dissection were respectively 5.44±2.63,2.92±1.87,10.81±4.78 in the robotic group and 3.11±1.82,1.62±1.33,7.76±3.34 in the laparoscopic group,with statistically significant differences between groups (t =3.340,2.689,2.522,P＜0.05).(3) Follow-up and survival situations:of 162 patients,148 were followed up for 2-14 months,with a median time of 8 months.During the follow-up,patients in the 2 groups had tumor-free survival.Conclusions Da Vinci robotic surgical system-assisted radical gastrectomy is safe and feasible.Compared with laparoscopy-assisted radical gastrectomy for locally advanced GC,it has advantages of clear vision of the local anatomy,less intraoperative bleeding,more numbers of lymph nodes dissected in the upper region of pancreas and lighter pancreatic injure,meanwhile,it has also certain operating advantages around the great vessels and in the deep and narrow spaces.

Objective To observe the effect of Shenfu injection on lung injury caused by hemor-rhagic shock(HS)in rats and explore the related potential mechanism.Methods Thirty-six SPF healthy male SD rats,aged 16-17 weeks,weighing 400-600 g,were randomly divided into three groups:sham op-eration group(group SH),HS group(group HS),and Shenfu injection group(group SF),12 rats in each group.In group SH,only the right femoral vein and femoral artery were separated after anesthesia,and ve-nous catheterization was not performed.HS model was established in groups SF and HS.In group HS,liquid resuscitation was performed through an intravenous catheter,and the resuscitation fluid consisted of the auto-blood lost and the compound sodium chloride injection equivalent to 1.5 times the blood loss and 10 ml/kg normal saline.In group SF,the resuscitation fluid consisted of the lost autoblood and the compound sodium chloride injection equivalent to 1.5 times the blood loss and Shenfu injection 10 ml/kg.The whole perfusion time was about 60 minutes.Six rats in the three groups were randomly anesthetized 24 and 48 hours after op-eration.The wet/dry weight ratio(W/D)of lung tissues was detected.The concentrations of interleukin-6(IL-6),IL-17,IL-10,and transforming growth factor-β(TGF-β)were detected by ELISA,the mRNA ex-pression of retinoic acid-related orphan nuclear receptor γt(RORγt),transcription factor forkhead box pro-tein 3(Foxp3),and hypoxia-inducible factor-1α(HIF-1α)in lung tissues were detected by PCR.The pro-tein contents of RORγt,Foxp3,HIF-1α,aquaporin 1(AQP1),and AQP5 in lung tissue were detected by Western blot.Pathological changesunder HE staining light microscope and lung injury scores were observed.Results Compared with 24 hours after operation,W/D,the concentrations of IL-6 and IL-17,mRNA ex-pression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P＜0.05),the concentrations of IL-10,and TGF-β,Foxp3 mRNA expression and protein content,and AQP1 protein content were significantly increased in group SF 48 hours after operation(P＜0.05).Compared with group SH,W/D,the concentrations of IL-6,IL-17,IL-10,and TGF-β,mRNA expression and protein content of RORγt,Foxp 3,and HIF-1α,and lung injury score were significantly increased(P＜0.05),AQP1 and AQP5 protein contents were significantly decreased in groups HS and SF 24 and 48 hours after operation(P＜0.05),and alveolar structure was damaged under light microscope and alveolar interstitium was filled with a large amount of edematous fluid,during which a large number of inflammatory cells infiltra-ted.Compared with group HS,W/D,the concentrations of IL-6 and IL-17,mRNA expression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P＜0.05),the concen-trations of IL-10 and TGF-β,Foxp3 mRNA expression and protein content,AQP1 and AQP5 protein con-tents were significantly increased in group SF 24 and 48 hours after surgery(P＜0.05),and the alveolar structure was improved under light microscope,and edema was reduced,and the number of inflammatory cells was reduced.Conclusion Shenfu injection can regulate the balance between pro-inflammatory factors IL-6 and IL-17,and anti-inflammatory factors IL-10 and TGF-β,increase the protein content of AQP1 and AQP5 in lung tissue,and decrease the W/D and injury score in lung tissue,thus alleviating lung injury in HS rats.The mechanism may be related to the regulation of HIF-1α-RORγt/Foxp3 balance.

Objective To analyze the effects of Hual qi huang granules on children with mycoplasma pneumoniae pneumonia.Methods A randomized,multicenter parallel controlled clinical trial was carried out.A total of 3 000 cases of hospitalized children with mycoplasma pneumoniae pneumonia were selected.All of them were given treatment for mycoplasma pneumoniae pneumonia with macrolide antibiotics and symptomatic treatment.They were randomly divided into 2 groups:research group and control group.The children of research group were give oral Huai qi huang granules for three months.According to the classification of pneumonia,these two groups were divided into:lobar pneumonia research group,lobar pneumonia control group,lobular pneumonia research group,lobular pneumonia control group.The hospitalization duration of fever,length of hospital stay,the absorption area of lung inflammation and pneumonia severity sores were observed.The frequency of upper respiratory infections,bronchitis,pneumonia were observed in 3 months after discharge.Results 2 378 cases were investigated.The hospitalization duration of fever,length of hospital stay of research group were significantly shorter than that of in control group (P ＜ 0.001).The children with lobar pneumonia,2 weeks after treatment,the absorption of consolidation of the lobar pneumonia research group is significantly better than lobar pneumonia control group (P ＜0.001).After two weeks treatment,the pneumonia scores of lobar pneumonia research group is lower than lobar pneumonia control group (P ＜ 0.05).Followup of 3 months after hospital discharge,frequency of upper respiratory infection and bronchitis of research group,were significantly lower than that of control.In addition,appetite increased significantly in research group than control (P ＜ 0.001).There are 21 cases with drug associated adverse reactions (mild diarrhea),including 12 cases of research group,9 cases of control group,and there was no statistical significance (P ＞0.05).Conclusion Standard treatment combined with oral Huai qi huang granules in the treatment of mycoplasma pneumoniae pneumonia,can significantly shorten hospitalization duration of fever,length of hospital stay and reduce the severity score of pneumonia.Three months oral Huai qi huang granules can significant reduce the frequency of respiratory infections and bronchitis,also can increase patients appetite,and be safe.

Objective: To analyze the pathological features and their influence on the clinical outcome of non-nasopharyngeal EBV-associated carcinomas. Methods: One hundred and twenty cases of non-nasopharyngeal EBV-associated carcinoma confirmed by in situ hybridization were identified at Zhejiang Cancer Hospital from January 1, 2006 to May 1, 2018, and the clinicopathological data were collected and analyzed using Kaplan-Meier survival analysis, Cox univariate and multivariate analysis. Results: One hundred and twenty cases were involved in the study; the male to female ratio was 1∶1; patients′ age range was 24 to 89 years (median 50 years). The primary sites were large parotid glands (62 cases), lung(26 cases), stomach(15 cases), and others (oral, oropharynx, larynx, cervix, liver; totally 17cases). Non-nasopharyngeal EBV-associated cancer could be divided into two histological types according to the amount of interstitial lymphocytes: type Ⅰ was "lymphoepithelial-like carcinoma" and rich in stromal lymphocytes; type Ⅱ lacked lymphocytic infiltration. Ninety-eight primary tumor samples could be classified morphologically: 43 cases were as type Ⅰ and 55 cases as typeⅡ; the distribution of type Ⅰ was 57.4% (27/47) in large parotid glands, 20.8% (5/24) in lung, 4/13 in stomach, and 7/14 in other sites. Complete treatment and survival data were obtained for 114 patients. According to the TNM staging criteria of WHO, 52 patients were at early stages (Ⅰ-Ⅱ) and 62 were at advanced stages (Ⅲ-Ⅳ); 102 patients underwent surgery. Seventy-four patients received adjuvant chemotherapy before or after surgery, and 52 patients received local radiotherapy. Kaplan-Meier survival analysis showed that patients with type Ⅱ EBV-associated carcinoma had a worse prognosis than patients with type Ⅰtumors (P=0.010 2). In addition, vascular invasion(P=0.021 8),neural recidivism(P=0.000 1),advanced stage(P=0.017 1),lymph node metastasis (P=0.005 0) and chemotherapy (P=0.013 2) were poor prognostic factors; female patients had better survival than male (P=0.028 4). Cox multivariate regression analysis found that lymph node metastasis (95%CI: 1.489-13.830, P=0.007 6) and neural recidivism (95%CI: 1.228-6.544, P=0.014 7) were independent adverse prognostic factors. Cox multivariate regression analysis after stratification by site revealed that radiotherapy was a preferable prognostic factor for EBV-associated carcinoma of the large salivary glands (95%CI: 0.003-0.569, P=0.016 8). Conclusion EBV associated carcinoma can be divided into two types, for which type Ⅰ was with abundant interstitial lymphocytes and type Ⅱ was lack of interstitial lymphocytes. TypeⅡ EBV-associated carcinoma has a worse prognosis than type Ⅰ. Radiation therapy can prolong the survival time of patients with primary EBV-associated carcinoma of large salivary glands.

Objective: To investigate the incidence of postoperative pancreatic fistula (POPF) and its risk factors after radical gastrectomy. Methods: The prospective study was conducted. The clinicopathological data of 2 089 patients who underwent radical gastrectomy in 22 medical centers between December 2017 and November 2018 were collected, including 380 in the Zhongshan Hospital of Fudan University, 351 in the Renji Hospital of Shanghai Jiaotong University School of Medicine, 130 in the Ruijin Hospital of Shanghai Jiaotong University School of Medicine, 139 in the Peking University Cancer Hospital, 128 in the Fujian Provincial Cancer Hospital, 114 in the First Hospital Affiliated to Army Medical University, 104 in the First Affiliated Hospital of Nanchang University, 104 in the Affiliated Hospital of Qinghai University, 103 in the Weifang People′s Hospital, 102 in the Fujian Medical University Union Hospital, 99 in the First Affiliated Hospital of Air Force Medical University, 97 in the Sir Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine, 60 in the Hangzhou First People′s Hospital Affiliated to Zhejiang University School of Medicine, 48 in the Fudan University Shanghai Cancer Center, 29 in the First Affiliated Hospital of Xi′an Jiaotong University, 26 in the Lishui Municipal Central Hospital, 26 in the Guangdong Provincial People′s Hospital, 23 in the Jiangsu Province Hospital, 13 in the First Affiliated Hospital of Sun Yat-Sen University, 7 in the Second Hospital of Jilin University, 4 in the First Affiliated Hospital of Xinjiang Medical University, 2 in the Beijing Chao-Yang Hospital of Capital Medical University. Observation indicators: (1) the incidence of POPF after radical gastrectomy; (2) treatment of grade B POPF after radical gastrectomy; (3) analysis of clinicopathological data; (4) analysis of surgical data; (5) risk factors for grade B POPF after radical gastrectomy. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using ANOVA. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Univariate analysis was conducted using the t test or chi-square test based on data excluding missing data of clinico-pathological and surgical data. Multivariate analysis was conducted using the Logistic regression model based on factors with P<0.20 in univariate analysis. Results: There were 2 089 patients screened for eligibility, including 1 512 males, 576 females and 1 without sex information, aged (62±11)years. The body mass index (BMI) was (23±3)kg/m². (1) The incidence of POPF after radical gastrectomy: the total incidence rate of POPF in the 2 089 patients was 20.728%(433/2 089). The incidence rates of biochemical fistula, grade B pancreatic fistula, and grade C pancreatic fistula were 19.627%(410/2 089), 1.101%(23/2 089), 0, respectively. (2) Treatment of grade B POPF after radical gastrectomy: 2 of 23 patients with grade B POPF after radical gastrectomy had drainage tube placed for more than 21 days and received anti-infective therapy. Four of 23 patients with grade B POPF after radical gastrectomy had ascites detected by imaging examination, of which 2 received peritoneal drainage guided by ultrasound, 1 received failed puncture drainage, 1 received no puncture drainage, and they were given anti-infective therapy. Eleven of 23 patients with grade B POPF after radical gastrectomy had no ascites detected by imaging examinations, and they were given anti-infective therapy and inhibitors of pancreas secretion for clinical manifestation as fever or elevated white blood cells. Six patients with no typical clinical manifestations were given somatostatin to inhibite pancreas secretion and prolonged duration of abdominal drainage tube placement (with a median time of 7 days). All the 23 patients recovered well after treatment, without reoperation. (3) Analysis of clinicopathological data: for the 2 089 patients, BMI, cases with or without neoadjuvant therapy were (23±3)kg/m^2, 1 487, 160 of patients without pancreatic fistula, (23±3)kg/m^2, 386, 22 of patients with biochemical fistula, and (24±3)kg/m^2, 22, 1 of patents with grade B pancreatic fistula, showing significant differences between the three groups (F=5.787, χ²=8.269, P<0.05). (4) Analysis of surgical data: for the 2 089 patients, cases with open surgery, laparoscopic assisted surgery, totally laparoscopic surgery (surgical method), cases with D₁ lymph lode dissection, D₂ lymph lode dissection, and other lymph lode dissection (range of lymph lode dissection), cases with no omentectomy, partial omentectomy, and total omentectomy (range of omentectomy), cases with no usage of energy facility, usage of CUSA, LigaSure, LigaSure+ CUSA as energy facility, cases with or without biological glue, the number of lymph node dissection were 737, 624, 292, 24, 1 580, 51, 418, 834, 381, 63, 1 530, 23, 16, 1 431, 201, 33±14 of patients without pancreatic fistula, 146, 189, 74, 11, 389, 9, 110, 171, 128, 35, 359, 6, 9, 378, 31, 31±14 of patients with biochemical fistula, and 14, 5, 4, 0, 20, 3, 6, 13, 4, 2, 18, 1, 2, 22, 1, 37±16 of patients with grade B pancreatic fistula, showing significant differences between the three groups (χ²=15.578, 9.397, 15.023, 28.245, 8.359, F=4.945, P<0.05). (5) Risk factors for grade B POPF after radical gastrectomy: results of univariate analysis showed that usage of energy facility was a related factor for grade B POPF after radical gastrectomy (χ²=9.914, P<0.05). Results of multivariate analysis showed that laparoscopic assisted surgery, combined evisceration, application of LigaSure + CUSA, the number of lymph lode dissection were independent factors for for grade B POPF after radical gastrectomy (odds ratio=0.168, 3.922, 9.250, 1.030, 95% confidence interval: 0.036-0.789, 1.031-14.919, 1.036-82.602, 1.001-1.059, P<0.05). Conclusions The incidence of grade B POPF after radical gastrectomy is relatively low. Laparoscopic assisted surgery, combined evisceration, application of LigaSure + CUSA, and the number of lymph lode dissection are independent risk factors for grade B POPF. Trial Registration: This study was registrated at ClinicalTrial.gov in United States with the registration number of NCT03391687.