Objective:To analyze the drug-resistant gene loci of Mycoplasma pneumoniae (MP) using metagenomic next-generation sequencing (mNGS). Methods:From November 2022 to October 2023, 697 clinical samples (including sputum, alveolar lavage fluid and blood) of 686 children with Mycoplasma pneumoniae positive detected by mNGS were retrospectively analyzed. Samples were divided into intensive care unit (ICU) group and non-ICU group, Chi-square test was used to compare groups, and Mann-Kendall trend test was used to analyze the change trend of the detection rate of drug resistance gene loci over time. Results:Of the 697 samples, 164 were from the ICU group and 533 were from the non-ICU group. The detection rate of Mycoplasma pneumoniae resistance gene was 44.3% (309/697), and all detected drug-resistant gene loci of MP were A2063G. The detection rate of Mycoplasma pneumoniae in ICU group was 50.0% (82/164), and the detection rates of Mycoplasma pneumoniae resistance gene loci in sputum, alveolus lavage fluid and blood samples were 75.0% (18/24) and 48.4% (62/128), respectively. The detection rate in sputum was higher than alveolus lavage fluid samples ( χ2=5.72, P=0.017). The detection rate of Mycoplasma pneumoniae in non-ICU group was 42.6% (227/533), the detection rate of Mycoplasma pneumoniae resistance gene loci in sputum and alveolar lavage fluid was 40.0% (16/40), 44.3% (201/454), and no detection rate in blood samples (0/12). There was no significant difference in the detection rate of alveolar lavage fluid and sputum ( χ2=0.27, P=0.602). From November 2022 to October 2023, the detection rate of submitted samples showed an increasing trend month by month (overall: Z=3.99, ICU inspection group: Z=2.93, non-ICU group: Z=3.01, all P<0.01). Among the bacteria commonly detected with Mycoplasma pneumoniae, Streptococcus pneumoniae accounted for the highest proportion, the detection rate was 15.5% (108/697), and Epstein-Barr virus accounted for the highest proportion of 17.6% (123/697). Conclusions:From November 2022 to October 2023, the detection rate of Mycoplasma pneumoniae drug resistance gene loci showed an increasing trend. The detection rate of drug resistance gene loci in sputum samples of ICU group was higher than alveolus lavage fluid. No new drug resistance site were detected.

Bluetongue virus is an arbovirus that seriously harms ruminants such as sheep,this study aims to investigate the molecular mechanism of bluetongue virus infection and host cell interferon antiviral immune response.The study was conducted to characterize the mRNA expression of inter-feron pathway genes by real-time fluorescence quantitative PCR,as well as Western blot analysis of MDA5,TRAF3,RIG-Ⅰ,and TBK1 protein expression in BHK-21 cells induced by BTV with a multiplicity of infections(MOI)of 1 for 18,24,and 36 h.The results showed that the most pro-nounced changes in the expression of interferon signaling pathway genes were observed at 24 h of induction,the gene mRNA expression levels of the IFN-α,IFN-β,RIG-Ⅰ,TBK1,MDA5,VISA,and TRAF3 genes were upregulated.However,the mRNA expression levels of IKKε and TRAF6 genes were downregulated.At the protein level,MDA5 and TBK1 proteins were upregulated while RIG-1 and TRAF3 proteins were downregulated,which showed that BTV infection induces a typeⅠ interferon immune response in BHK-21 cells.This study lays the foundation for further exploring the antiviral immunity mechanism of IFN-Ⅰ signaling pathway regulatory genes in host cells infected with BTV infection.

Humans ; Multiple Pulmonary Nodules ; Lung ; Lung Neoplasms/surgery* ; Solitary Pulmonary Nodule/surgery* ; Bronchoscopy

The role of glial scar after intracerebral hemorrhage(ICH)remains unclear.This study aimed to inves-tigate whether microglia-astrocyte interaction affects glial scar formation and explore the specific function of glial scar.We used a pharmacologic approach to induce microglial depletion during different ICH stages and examine how ablating microglia affects astrocytic scar formation.Spatial transcriptomics(ST)analysis was performed to explore the potential ligand-receptor pair in the modulation of microglia-astrocyte interaction and to verify the functional changes of astrocytic scars at different periods.During the early stage,sustained microglial depletion induced disorganized astrocytic scar,enhanced neutrophil infiltration,and impaired tissue repair.ST analysis indicated that microglia-derived insulin like growth factor 1(IGF1)modulated astrocytic scar formation via mechanistic target of rapamycin(mTOR)signaling activation.Moreover,repopulating microglia(RM)more strongly activated mTOR signaling,facilitating a more protective scar formation.The combination of IGF1 and osteopontin(OPN)was necessary and sufficient for RM function,rather than IGF1 or OPN alone.At the chronic stage of ICH,the overall net effect of astrocytic scar changed from protective to destructive and delayed microglial depletion could partly reverse this.The vital insight gleaned from our data is that sustained microglial depletion may not be a reasonable treatment strategy for early-stage ICH.Inversely,early-stage IGF1/OPN treatment combined with late-stage PLX3397 treatment is a promising therapeutic strategy.This prompts us to consider the complex temporal dynamics and overall net effect of microglia and astrocytes,and develop elaborate treatment strategies at precise time points after ICH.

Objective:To investigate the clinical value of stylized laparoscopic hemicolec-tomy for left colon cancer.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 174 patients who underwent laparoscopic hemicolectomy for left colon cancer in the First Affiliated Hospital of Nanjing Medical University from January 2016 to December 2020 were collected. There were 106 males and 68 females, aged 59(range, 17?86)years. All patients underwent stylized laparoscopic hemicolectomy for left colon cancer. Observation indicators: (1) surgical situations; (2) postoperative complications; (3) postoperative histopathological examinations; (4) follow-up. Follow-up was conducted using outpatient examination or telephone interview to detect tumor recurrence and metastasis and survival of patients up to October 2021. Follow-up was performed once every 3 months within postoperative 2 years, once every 6 months within postoperative 2 to 5 years and once a year after postoperative 5 years, with the end point as tumor recurrence and metastasis or death of patients. Measurement data with normal distribution were represented as Mean±SD and measurement data with skewed distribution were represented as M(range) or M( Q1, Q3). Count data were described as absolute numbers or percentages. Kaplan-Meier method was used to draw survival curve and calculate survival rate. Results:(1) Surgical situations. All the 174 patients underwent stylized laparoscopic hemicolectomy for left colon cancer successfully, including 6 cases receiving preoperative enteral stent placement due to bowel obstruc-tion, 3 cases receiving defunctioning ileostomy and 1 case receiving Hartmann procedure. The operation time, volume of intraoperative blood loss, time to postoperative initial defecation and duration of postoperative hospital stay of the 174 patients were 97(80,106)minutes, 45(25,60)mL, 5(3,6)days and 7(6,8)days, respectively. (2) Postoperative complications. Twelve of the 174 patients had complications, including 4 cases with incision infection or fat liquefaction, 3 cases with anastomotic leakage, 2 cases with incomplete bowel obstruction, 1 case with abdominal hemo-rrhage, 1 case with chylous leakage and 1 case with pulmonary infection. The 2 cases with anastomotic leakage underwent ileostomy. The patient with abdominal hemorrhage underwent laparotomy to stop bleeding. One elder patient died of postoperative pulmonary infection. The other patients with complications recovered with conservative treatment. (3) Postoperative histopatho-logical examinations. Of the 174 patients, there were 27 cases in stage Ⅰ of TNM staging, 68 cases in stage Ⅱ, 77 cases in stage Ⅲ and 2 cases in stage Ⅳ. There were 9 cases with well differentiated tumor, 107 cases with moderately differentiated tumor and 58 cases with poorly differentiated tumor. The number of lymph node detected, the number of positive lymph node and tumor diameter of the 174 patients were 19(15,23), 0(0,2) and 4(3,5)cm, respectively. Of the 174 patients, there were 79 cases with lymph node metastases, 21 cases with cancerous nodules, 35 cases with vascular invasion and 29 cases with nerve invasion. (4) Follow-up. Of the 174 patients, 157 cases were followed up for 27(range, 1?70)months. Of the 157 patients who conducted follow-up, 20 cases had tumor metastasis, including 9 cases with multiple metastasis, 5 cases with liver metastasis, 4 cases with lung metastasis, 1 case with bone metastasis and 1 case with spleen metastasis. The 5-year overall survival rate and tumor free survival rate of the 157 patients were 90.9% and 80.8%, respectively.Conclusion:The stylized laparoscopic hemicolectomy for left colon cancer is safe and feasible.

OBJECTIVE：To study the composition and contents of flavonoids chemical components in waste material during industrialization of Pueraria thomsonii resources，and to provide reference for comprehensive development and reasonable utilization of the variety. METHODS ：Using“No. 2 Gange”of P. thomsonii from Jiangxi as objects ，UPLC-Q-TOF-MS and HPLC method were adopted to detect the components and contents of flavonoids in the root （with or without cortex ），cortex，flower， fibrous root ，stem，head and dregs （with or without cortex ）of P. thomsonii as well as dry matter of industrial wastewater （with or without cortex ）after precipitation of pueraria powder. RESULTS ：The linearity ，precision，repeatability，stability and recovery of the established method for content determination of 7 flavonoids（puerarin，daidzin，iridoxine-7-O-xylose glucoside ，genistin， iridin，daidzein and kakkalide ）were all in line with the requirements. Totally 12 kinds of flavonoids were identified ，among which the flavonoids in the root ，cortex，stem，fibrous root ，head and dregs of P. thomsonii as well as dry matter of industrial wastewater were the same ，mainly were puerarin ，daidzin，genistein，daidzein and malonyl-daidzein. The flower of P. thomsonii mainly included iridoxine- 7-O-xylose glucoside ，genistin，iridin，kakkalide，6″-O-xylosyldaidzein，but the components as puerarin ， daidzin and its aglycone were not be detected. The content of puerarin in the head of P. thomsonii was the highest （5.765％）. The contents of puerarin in root and dregs of P. thomsonii as well as dry matter of industrial waste-water in samples with cortex were all higher than in corresponding peeled sample. CONCLUSIONS ：The waste material from the industrialization of P. thomsonii resources contains a lot of flavonoids with rich species and high content ，and can be used as an important raw material for obtaining flavonoids such as puerarin.

OBJECTIVE： To establish the method for simultaneous determination of four known related substances （olmesartan，olmesartan ester dimer ，olmesartan ester alkene ，benzothiadiazine impurity ，called impurity A ，B，C，D for short ）in Olmesartan medoxomil hydrochlorothiazide tablets. METHODS ：HPLC-principal component self-control with correction factor were adopted. The determination was performed on YMC-Triart C 8 column with mobile phase A consisted of acetonitrile- 0.015 mol/L potassium dihydrogen phosphate solution （pH adjusted to 2.8 with phosphoric acid ）（70 ∶ 30，V/V），mobile phase B consisted of acetonitrile-0.015 mol/L potassium dihydrogen phosphate solution （pH adjusted to 2.8 with phosphoric acid ）（15 ∶ 85，V/V）at the flow rate of 0.8 mL/min（gradient elution ）. The detection wavelength was set at 265 nm，and column temperature was 25 ℃. The temperature of injector was 4 ℃；the injection volume was 10 μL. RESULTS：The correction factors of impurity A ，B，C，D were 1.42，1.17，0.89，0.92，respectively. The linear range of olmesartan medoxomil ，hydrochlorothiazide and impurity A ，B，C，D were 0.252 7-7.580 0，1.152 1-4.562 9，0.244 0-18.299 0，0.244 7-3.670 8，0.265 2-3.978 3 and 0.149 9-4.497 3 μg/mL（r≥ 0.999 7），respectively. The limits of detection were 0.084 2，0.050 7，0.081 3，0.081 6，0.088 4，0.050 0 μg/mL，respectively. The quantitative limits were 0.252 7，0.152 1，0.244 0，0.244 7，0.265 2 and 0.149 9 μg/mL，respectively. The results of intermediate precision ，stability（24 h）and repeatability tests all met the relevant requirements. The average recovery rates were 104.00％-108.04％，102.00％-104.94％，100.99％-106.89％，92.00％-95.18％，102.00％-105.06％，103.90％-107.00％（n＝3）， respectively. The contents of impurity A ，B and D in 3 batches of Olmesartan medoxomil hydrochlorothiazide tablets were 0.90％ -1.00％ ，0-0.11％ ，0.16％ -0.24％ ，respectively. The impurity C and other impurities were not detected. There is no significant difference between the results measured by the established method and by the external standard method. CONCLUSIONS：The method has been proved to be highly sensitive and reproducible. It can be used to simultaneously determine four known substances in Olmesartan medoxomil hydrochlorothiazide tablets.

Objective To investigate the application value of superior mesenteric artery (SMA)-oriented complete mesocolic excision (CME) in the treatment of right colon cancer.Methods The retrospective cohort study was conducted.The clinicopathological data of 955 patients with right colon cancer who were admitted to the First Affiliated Hospital of Nanjing Medical University from January 2013 to June 2018 were collected.There were 514 males and 441 females,aged from 18 to 96 years,with a median age of 65 years.Of the 955 patients,377 undergoing SMA-oriented CME of right colon with the lymph node dissection along the left boundary of SMA were allocated into SMA-oriented group,and 578 undergoing superior mesenteric vein (SMV)-oriented CME of right colon with the lymph node dissection along the left boundary of SMV were allocated into SMV-oriented group.Observation indicators:(1) intraoperative and postoperative conditions;(2) postoperative complications;(3) postoperative pathological examinations;(4) follow-up and survival situations.Follow-up was performed by telephone interview and outpatient examination once every 3-6 months within 2 years after surgery and once a year after 2 years up to January 2019,using tumor recurrence and metastasis or death as the end point.Follow-up included physical examination and tumor marker test,including carcino embryonic antigen,CA19-9,chest and abdomen CT examination and enteroscopy.Measurement data with skewed distribution were described as M (P25,P75),and comparison between groups was done using the Mann-Whitney U test.Count data were described as absolute numbers or percentages,and comparison between groups was analyzed using the chi square test.Ordinal data were analyzed using the rank sum test.Kaplan-Meier method was used to calculate survival time and rate,and draw survival curve.Log-rank test was used for survival analysis.Patients with loss to follow-up were involved in survival analysis as censored data.Results (1) Intraoperative and postoperative conditions:the operation time,volume of intraoperative blood loss,duration of postoperative hospital stay were 100 minutes (90 minutes,110 minutes),50.0 mL (50.0 mL,70.0 mL),8 days (8 days,10 days) in the SMA-oriented group,and 110 minutes (90 minutes,135 minutes),50.0 mL (50.0 mL,122.5 mL),10 days (8 days,12 days) in the SMV-oriented group,showing significant differences between the two groups (Z=-5.400,-5.799,-7.461,P＜0.05).After the exclusion of 47 patients unsuitable for defecation analysis because of postoperative complications,365 in the SMA-oriented group and 543 in the SMV-oriented group were analyzed.The time to first defecation,the maximum number and the median number of daily defecation postoperatively were 5 days (3 days,5 days),2.0 (1.0,2.5),1.0 (1.0,1.0) in the SMA-oriented group,which showed no significant difference from 4 days (3 days,5 days),2.0 (1.0,3.0),1.0 (1.0,1.0) in the SMV-oriented group (Z=-1.622,-1.541,-1.024,P＞ 0.05).(2) Postoperative complications:cases with postoperative complications,cases with incisional liquefaction or infection,cases with anastomostic leakage,cases with delayed gastric emptying,cases with intra-abdominal bleeding,cases with complete or incomplete ileus,cases with anastomostic bleeding,cases with intra-abdominal infection,cases with disruption of wound,the number of death were 55,10,3,3,2,2,1,1,1,1 in the SMA-oriented group,which showed no significant difference from 83,30,13,4,3,8,3,6,2,3 in the SMV-oriented group,respectively (x2 =0.045,3.662,2.926,0.034,0.001,1.604,0.352,1.873,0.048,0.352,P＞0.05).There were 32 of 377 patients in the SMA-oriented group and 14 of 578 in the SMV-oriented group with chylous leakage,showing a significant difference between the two groups (x2 =18.312,P＜ 0.05).Patients with chylous leakage were improved after conservative treatment,without reoperation.Patients with other complications were improved after anti-infection,fluid infusion,and reoperation.Four of 955 patients died after surgery.(3) Postoperative pathological examinations:patients with stage Ⅰ,stage Ⅱ,and stage Ⅲ (pathological TNM staging),patients with high-differentiation,mid-differentiation,and low-differentiation (tumor differentiation degree),length of intestine specimen,number of positive lymph nodes,maximum tumor diameter,patients with cancer nodules,patients with vascular invasion,patients with perineural invasion were 57,174,146,30,174,173,23 cm (21 cm,26 cm),0 (0,2),5.0 cm (3.0 cm,6.0 cm),37,81,53 in the SMA-oriented group,which showed no difference from 66,280,232,33,303,242,23 cm (21 cm,25 cm),0 (0,2),5.0 cm (3.5 cm,6.0 cm),80,108,82 in the SMV-oriented group (Z=-1.020,-0.216,-0.243,-0.220,-0.814,x2=3.441,1.127,0.003,P＞0.05).The number of harvested lymph nodes was 22.0 (17.0,27.0) and 18.0 (15.0,22.0) in the SMA-oriented group and SMV-oriented group,respectively,with a significant difference between the two groups (Z=-7.800,P＜0.05).There were 202 patients extracted for further analysis.The number of harvested lymph nodes and harvested central lymph nodes was 25.0 (20.0,31.3),5.0 (3.0,8.0) of 166 patients in the SMA-oriented group,and 21.5 (18.0,28.8),1.5 (0,4.5) of 36 patients in the SMV-oriented group,respectively,showing significant differences between the two groups (Z =-1.995,-4.309,P＜0.05).(4) Follow-up and survival situations:840 of 955 patients including 346 in the SMA-oriented group and 494 in the SMV-oriented group were followed up for 1.0-73.2 months,with a median time of 31.5 months.SMA-oriented group had a higher 5-year overall survival rate than SMV-oriented group (91.8％ vs.84.9％,x2 =4.384,P＜0.05),but had no significant difference in the 5-year tumor-free survival rate compared with the SMV-oriented group (84.4％ vs.78.2％,x2=2.158,P＞0.05).Conclusion Compared with SMV-oriented CME of right colon,SMA-oriented CME of right colon is safe and feasible,with larger number of harvested lymph nodes,which can achieve complete lymph node dissection.

Objective To analyze the risk factors for anastomotic leakage after low anterior resection(LAR) for rectal cancer.Methods The retrospective study of 1 336 patients with rectal cancer who underwent LAR from Jan.2013 to Dec.2017 in the Department of Colorectal Surgery,First Affiliated Hospital of Nanjing Medical University was conducted.There were 838 male and 498 female patients.The median age was 62 (53,70) years.Observation indicators:①intraoperative conditions,including the performance of preventive enterostomy,the preservation of left colic artery,the placement of transanal decompression tube;②postoperative conditions,for example,postoperative clinical symptoms and physical examination,postoperative hospital stay.Anastomotic leakage patients were graded according to the grading system of anastomotic leakage following anterior resection of the rectum proposed by the International Study Group of Rectal Cancer in 2010.Measurement data with non-normal distribution were described as median,and comparison between groups was done using Mann-Whitney U test.Ranked data were compared with rank sum test.Comparison of count data between groups were analyzed using the Chi-square test or Fisher exact test.Potential risk factors for anastomotic leakage were further analyzed with multiple logistic regression.Results Anastomotic leakage was noted in 138 (10.4％) of 1 336 patients with 93 patients (7.0％) of Grade B and 46 patients (3.4％) of Grade C respectively.The occurrence of Grade C anastomotic leakage was on the 3th day postoperatively,which was earlier than Grade B anastomotic leakage [5 (3,7) d,Z =2.746,P =0.006].There were significant differences in sex,BMI,placement of transanal decompression tube and tumor location between anastomotic leakage cases and non-anastomotic leakage cases by Chi-squared test (x2 =10.897,9.033,6.343,6.34,Z =-2.380,P ＜0.05).On multiple logistic regression analysis,male sex (OR =1.840,95 ％ CI:1.210-2.799,P =0.004),BMI ≥ 24 kg/m2 (OR =1.573,95 ％ CI:1.088-2.274,P =0.016),placement of transanal decompression tube(OR =2.418,95％ CI:1.196-4.888,P =0.014),middle rectal cancer (OR =1.536,95％ CI:O.836-2.822,P =0.167),low rectal cancer(OR =1.989,95％ CI:1.068-2.822,P =0.03),ultralow rectal cancer (OR =2.908,95％ CI:1.289-6.560,P =0.01) were independent risk factors of anastomotic leakage.Conclusion Male sex,high BMI,placement of transanal decompression tube and low rectal tumor were independent risk factors of anastomotic leakage for patients receiving LAR.