Objective To investigate the prognostic value of C5L2 in patients with hepatocellular carcinoma (HCC).Methods The data of 175 patients with HCC who underwent curative resection at Zhongshan Hospital,Fudan University from Oct.,2012 to Sep.,2013 were analyzed retrospectively.The correlation between C5L2 and clinicopathologic characteristics were explored.COX regression model was used to determine the influence of clinical parameters on predicting recurrence,and the prognostic value of combined application of C5L2 and AFP were evaluated by Kaplan-Meier method.In vitro,the expression of C5L2 were tested in 5 HCC cell lines,and Hep3B and Huh7 were chosen for down-regulation and up-regulation of C5L2,respectively,the abilities of invasion and migration were examined by transwell and the potential mechanism was explored.Results The C5L2 expression was correlated to gender,tumor size and recurrence,and the recurrence rate of low C5L2 expression group was higher.Also,the multivariate analysis showed that C5L2 low expression was an independent risk factor for recurrence.Moreover,the combined application of C5L2 and AFP could estimate prognosis more effectively.Knockdown of C5L2 in Hep3B promoted the invasiveness and motility,and increased the level of β-catenin and MMP2;conversely,overexpression of C5L2 in Huh7 inhibited the invasiveness and motility,and decreased the level of β catenin and MMP2.Conclusions C5L2 could be regarded as an auxiliary indicator for prognosis of HCC,thereby the evaluation of C5L2 could help with making effective and comprehensive management for HCC patients.

Objective:To verify the performance of the next-generation sequencing (NGS) platform and evaluate the application of NGS, droplet digital PCR (ddPCR) and super amplification refractory mutation system (super-ARMS) in the detection of circulating free DNA (cfDNA) mutations in patients with non-small-cell lung cancer (NSCLC).Methods:A total of 75 patients with NSCLC in the respiratory department of Zhongshan Hospital Affiliated to Fudan University were enrolled. The standards, cfDNA from 25 patients with newly diagnosed and untreated NSCLC, and self-made mixed samples mixed with hemoglobin (1 000 mg /dl), bilirubin (500 mg/l), fat emulsion (2%), enterococcus gDNA and Escherichia coli gDNA were used to verify the blank limit, analytical sensitivity, precision, accuracy and specificity of NGS platform. The cfDNA mutations of 75 NSCLC patients were detected by ddPCR and NGS, and the mutation positive rates of the two platforms were compared. The linear relationship between the two platforms was compared by Pearson correlation test. 12 patients were selected by simple random sampling for the detection of plasma super-ARMS platform. The performance of three platforms in the detection of plasma cfDNA mutation in patients with NSCLC was compared.Results:The blank limit of NGS platform was set to 0.00%, the analytical sensitivity was 0.2%, the intra-assay precision and inter-assay precision were 100%. The test results were not affected by endogenous hemoglobin, bilirubin or fat emulsion in plasma or exogenous DNA interference, and the analysis specificity was good. The mutation positive rates of plasma cfDNA in 75 NSCLC patients detected by ddPCR and NGS were 61.33% and 60.00%, respectively. The complete coincidence rate was 89.33%, which suggests there was a positive correlation between the mutation abundance of NGS and ddPCR ( r=0.984, P=0.001). Among the plasma of 12 NSCLC patients, the results of NGS, ddPCR and super-ARMS were completely consistent in 7 cases, including 2 wild-types and 5 mutants. Conclusion:The NGS platform was verified to be useful for cfDNA mutation detection in patients with NSCLC. The ddPCR, NGS and super-ARMS have their own advantages in detecting cfDNA mutations in patients with NSCLC.

OBJECTIVE: To investigate the effects of gene of phosphate and tension homology (PTEN)-induced putative kinase 1 (PINK1)/Parkin pathway on hippocampal mitophagy and cognitive function in mice with sepsis-associated encephalopathy (SAE) and its possible mechanism. METHODS: A total of 80 male C57BL/6J mice were randomly divided into Sham group, cecal ligation puncture (CLP) group, PINK1 plasmid transfection pretreatment groups (p-PINK1+Sham group, p-PINK1+CLP group), empty vector plasmid transfection control group (p-vector+CLP group), with 16 mice in each group. The mice in CLP groups were treated with CLP to reproduce SAE models. The mice in the Sham groups were performed laparotomy only. Animals in the p-PINK1+Sham and p-PINK1+CLP groups were transfected with PINK1 plasmid through the lateral ventricle at 24 hours before surgery, while mice in the p-vector+CLP group were transfected with the empty plasmid. Morris water maze experiment was performed 7 days after CLP. The hippocampal tissues were collected, the pathological changes were observed under a light microscope after hematoxylin-eosin (HE) staining, and the mitochondrial autophagy was observed under a transmission electron microscopy after uranyl acetate and lead citrate staining. The expressions of PINK1, Parkin, Beclin1, interleukins (IL-6, IL-1β) and microtubule-associated protein 1 light chain 3 (LC3) were detected by Western blotting. RESULTS: Compared with the Sham group, CLP group mice in Morris water maze experiment had longer escape latency, shorter target quadrant residence time, and fewer times of crossing the platform at 1-4 days. Under the light microscope, the hippocampal structure of the mouse was injured, the neuronal cells were arranged in disorder, and the nuclei were pyknotic. Under the electron microscope, the mitochondria appeared swollen, round, and wrapped by bilayer or multilayer membrane structures. Compared with the Sham group, CLP group had higher expressions of PINK1, Parkin, Beclin1, LC3II/LC3I ratio, IL-6 and IL-1β in hippocampus, indicating that sepsis induced by CLP could activated inflammatory response and caused PINK1/Parkin-mediated mitophagy. Compared with the CLP group, p-PINK1+CLP group had shorter escape latencies, spent more time in the target quadrant and had more number of crossings in the target quadrant at 1-4 days. Under the light microscope, the hippocampal structures of mice was destroyed, the neurons were arranged disorderly, and the nuclei were pyknotic. Under transmission electron microscope, swollen and rounded mitochondria and mitochondrial structure wrapped by double membrane or multilayer membrane structure were observed. Compared with the CLP group, the levels of PINK1, Parkin, Beclin1 and LC3II/LC3 ratio in the p-PINK1+CLP group were significantly increased [PINK1 protein (PINK1/β-actin): 1.95±0.17 vs. 1.74±0.15, Parkin protein (Parkin/β-actin): 2.06±0.11 vs. 1.78±0.12, Beclin1 protein (Beclin1/β-actin): 2.11±0.12 vs. 1.67±0.10, LC3II/LC3I ratio: 3.63±0.12 vs. 2.27±0.10, all P < 0.05], while the levels of IL-6 and IL-1β were significantly decreased [IL-6 protein (IL-6/β-actin): 1.69±0.09 vs. 2.00±0.11, IL-1β protein (IL-1β/β-actin): 1.11±0.12 vs. 1.65±0.12, both P < 0.05], suggesting that overexpression of PINK1 protein could further activate mitophagy and reduce the inflammatory response caused by sepsis. There was no statistically significant difference in the above pathological changes and related indicators between Sham group and p-PINK1+Sham group, CLP group and p-vector+CLP group. CONCLUSIONS PINK1 overexpression can further activate CLP-induced mitophagy by upregulating Parkin, thereby inhibiting inflammation response and alleviate cognitive function impairment in SAE mice.
Male ; Animals ; Mice ; Mice, Inbred C57BL ; Sepsis-Associated Encephalopathy ; Phosphates ; Actins ; Beclin-1 ; Interleukin-6 ; Autophagy ; Ubiquitin-Protein Ligases ; Cognitive Dysfunction ; Sepsis ; Mitochondria ; Protein Kinases

Objective:To investigate the effectiveness and feasibility of whole exome sequencing (WES), as a molecular diagnosis technique, for adult patients with genetic diseases.Methods:The present retrospective analysis included 445 adult patients (ages 18-80 years) with suspected genetic diseases who underwent whole exome sequencing (WES) from August 2021 to December 2022. The pathogenicity classification of each variant was assessed in accordance with the recommendations developed by the American Society of Medical Genetics and Genomics.Results:The overall positive rate of WES among adult patients with suspected genetic diseases was 28.08% (125/445). The highest positive rate was observed in the age group of 41-50 years (34.33%, 23/67). Among the diagnosed genetic diseases, those affecting the cardiovascular system (63.16%, 84/133), nervous system (18.05%, 24/133), and endocrine system (13.53%, 18/133) ranked as the top three. The most common genetic diseases identified through WES in adult patients were hypertrophic cardiomyopathy (18.80%, 25/133), dilated cardiomyopathy (16.54%, 22/133), Marfan syndrome (15.04%, 20/133), epilepsy (9.02%, 12/133), and familial hypercholesterolemia (4.51%, 6/133). The main causative genes identified included FBN1 (14.29%, 19/133), MYBPC3 (9.02%, 12/133), MYH7 (9.02%, 12/133), LDLR (3.76%, 5/133), TTN (3.76%, 5/133), and TNNI3 (3.01%, 4/133).Conclusion:Applying the WES technique in clinical practice can improve the diagnostic rate of adult genetic diseases, especially in adult patients with suspected genetic conditions involving the cardiovascular system, nervous system, and endocrine system.

Objective To observe the effects of transnasal humidified rapid-insufflation ventilatory exchange(THRIVE)on gastric air intake and SpO2 in patients with painless endoscopic mucosal resection(EMR)of colorectal polyps.Methods Seventy patients with painless colorectal polyps EMR,36 males,34 females,aged 18-64 years,BMI 18-25 kg/m2,ASA Ⅰ or Ⅱ.The patients were divided into two groups:high-flow oxygen inhalation group(group T)and conventional oxygen inhalation group(group C)by random number table method,35 cases in each group.Patients in group T were given THRIVE oxygen inhalation,and patients in group C were routinely inhaled oxygen through the nose.During anesthesia,the BIS value was maintained at 50-65.Bedside gastric ultrasonography was used to observe the gastric content and gastric air intake of patients.The HR and resting SpO2 in the supine position were recorded 5 minutes before the surgery(rest for 5 minutes),1 minute after entering the endoscope,when entering the endoscope to the ileocecal valve,and at the end of the surgery.The PaCO2,PaO2 and pH values were recorded 5 mi-nutes before surgery,when entering the endoscope to the ileocecal valve,and at the end of the surgery,so were gastric intake and satiety,and antral cross-sectional area(CSA)for 5 minutes before surgery and after surgery.The occurrence of mild,moderate,and severe hypoxia during anesthesia were recorded.The opera-tion time,the amount of propofol used during the operation,the number of dosage times of remifentanil,PACU residence time,and infusion during surgery,the occurrence of adverse reactions such as hypotension,hypertension,bradycardia,reflux aspiration,postoperative nausea and vomiting,and the sat-isfaction score of digestive endoscopy physicians and patients were recrded.Results Compared with group C,SpO2 in group T was significantly increased 1 minutes after entering the endoscope(P＜0.05),and PaCO2 was significantly decreased at the time of entering the endoscope to the ileocecal valve and the end of the surgery(P＜0.05),the incidence of mild and moderate hypoxia was significantly reduced in group T(P＜0.05).No positive gastric air intake and full stomach were found in both groups at rest for 5 minutes and after the surgery.There was no significant difference in the incidence of adverse reactions and patient satisfaction scores between the two groups.The satisfaction score of endoscopists in group T was significantly higher than that in group C(P＜0.05).Conclusion Compared with conventional nasal catheter oxygen inhalation,THRIVE ventilation does not cause hypercapnia or affect gastric intake and stomach content,does reduce the occurrence of mild to moderate hypoxia in patients undergoing painless EMR,does not in-crease the risk of reflux aspiration,and has a higher satisfaction of digestive endoscopy physicians.

Objective To evaluate the effect of endoscopic laryngeal mask on perioperative airway management and postoperative recovery in patients undergoing gastric endoscopic submucosal dissection(ESD).Methods Ninety patients,aged 18-64 years,BMI 18-25 kg/m2,ASA physical statusⅠorⅡ,who underwent elective gastric ESD were randomly divided into two groups:the endoscopic laryngeal mask group(group E)and the endotracheal tube group(group C),45 patients in each group.After induction of general anesthesia,group E received endoscopic laryngeal mask airway ventilation,and the endoscope was inserted through the endoscopic channel of the laryngeal mask,group C received tracheal intubation,and the endoscopy was inserted through the mouth.The successful time and one-time success rate of intubation,and the insertion time and withdrawal rate of endoscopy were recorded.The operative time,extubation time and PACU residence time were recorded.HR,MAP were recorded when the patient entered the room(T0),at the time of intubating(T1),inserting gastroscopy(T2),exiting gastroscopy(T3),extubation(T4),and leaving PACU(T5).The average airway pressure and peak airway pressure at T1-T3 were recorded.The airway sealing pressure and endoscopic view grading system(EVGS)grading of group E were recorded before and after changing the position,and at the end of surgery.The adverse reactions and the satisfaction of anesthesiologists and gastroenterologists were recorded.Results Compared with T0,HR and MAP were significantly increased at T1 and T4 between the two groups(P＜0.05).Compared with group C,the suc-cessful time of intubation,the extubation time,and PACU residence time were significantly shortened,HR and MAP were significantly decreased at T1 and T4,the incidence of choking cough during extubation,post-operative pharyngeal pain,and hoarseness were significantly decreased(P＜0.05).There were no signifi-cant differences in the one-time success rate of intubation,the insertion time and withdrawal rate of endosco-py between the two groups.Endoscopic laryngeal mask showed good sealing and alignment in group E.Conclusion Endoscopic laryngeal mask could shorten the success time of establishment of artificial airway in patients with gastric ESD,without interfering with digestive endoscopy operations,shorten extubation time and PACU retention time,maintain intraoperative hemodynamic stability,and reduce adverse reactions.

Objective:To analyze the efficacy of endoscopic laryngeal mask in obese patients undergoing upper gastrointestinal endoscopic mucosal resection (EMR).Methods:Adopting a prospective research approach, 90 obese patients who underwent upper gastrointestinal EMR from July 2020 to May 2022 in Nanjing First Hospital, Nanjing Medical University were selected. The patients were divided into endoscopic laryngeal mask group (using general anesthesia with endoscopic laryngeal mask ventilation) and nasal catheter group (using conventional non intubated intravenous anesthesia) by random digits table method with 45 cases in each. The EMR time; minimum pulse oxygen saturation (SpO 2) during perioperative period; changes of SpO 2, mean arterial pressure (MAP) and heart rate during perioperative period; palinesthesia time; postoperative postanesthesia care unit (PACU) residence time; adverse reactions; satisfaction levels of doctor and patient satisfaction were recorded. Results:There were no statistical difference in EMR time and postoperative PACU residence time between two groups ( P>0.05). There were no statistical difference in minimum SpO 2 preoperative and anesthesia induction period between two groups ( P>0.05); the minimum SpO 2 EMR period in endoscopic laryngeal mask group was significantly higher than those in nasal catheter group (0.990 ± 0.010 vs. 0.951 ± 0.037), and there was statistical difference ( P<0.01). There were no statistical difference in heart rate and MAP during perioperative period between two groups ( P>0.05). There were no statistical difference in SpO 2 entering the operating room and leaving the PACU between two groups ( P>0.05), SpO 2 immediately after endoscopic insertion and at the end of surgery in endoscopic laryngeal mask group was significantly higher than that in nasal catheter group (0.989 ± 0.009 vs. 0.976 ± 0.011 and 0.987 ± 0.010 vs. 0.981 ± 0.009), and there was statistical difference ( P<0.01). The palinesthesia time in endoscopic laryngeal mask group was significantly longer than that in nasal catheter group: (6.7 ± 1.1) min vs. (4.6 ± 1.2) min, and there was statistical difference ( P<0.01). Both groups did not experience aspiration, hoarseness or airway spasm. There were no statistical difference in the incidences of pharyngalgia, bradycardia, hypotension, abdominal pain and bloating, postoperative nausea and vomiting between two groups ( P>0.05). The incidence of bucking and body movement in endoscopic laryngeal mask group was significantly lower than that in nasal catheter group: 2.2% (1/45) vs. 24.4% (11/45), and there was statistical difference ( P<0.01). There were no statistical difference in the satisfaction level of patient between two groups ( P>0.05); the satisfaction levels of anesthesiologists and endoscopists in endoscopic laryngeal mask group were significantly higher than those in nasal catheter group: 95.6% (43/45) vs. 66.7% (30/45) and 88.9% (40/45) vs. 71.1% (32/45), and there were statistical differences ( P<0.01 or <0.05). Conclusions:The application of endoscopic laryngeal mask in upper gastrointestinal EMR in obese patients can effectively improve the hypoxia caused by insufficient ventilation. At the same time, it will not cause drastic fluctuations in hemodynamics, and it does not increase the incidence of throat discomfort, but it slightly extends the awakening time.

Objective To investigate the effect of nasal mask on preventing respiratory depression and hypoxia during anesthesia under painless gastroscopy.Methods 246 patients underwent elective painless gastroscopy from September 2022 to March 2023 were selected and divided into nasal catheter oxygen group(group C)and nasal mask oxygen group(group M)according to random number table method.The group C was inhaled oxygen by conventional nasal cannula,and in group M,oxygen was inhaled by a nasal mask.Patients in both groups were given 5 L/min preoxygenation for 3 min.After induction of anesthesia,gastroscopy was performed when modified observer's assessment of alertness/sedation(MOAA/S)score≤1.During anesthesia,hypoxia interventions were performed in group C based on percutaneous arterial oxygen saturation(SpO2)and in group M based on partial pressure of end-tidal carbon dioxide(PetCO2)and SpO2.The incidence of subclinical respiratory depression,hypoxia and severe hypoxia during anesthesia was recorded.The minimum SpO2 value during anesthesia was recorded;Implementation of intervention measures of jaw lifting,mask pressure oxygen and tracheal intubation.Record the number of gastroscope withdrawal cases.Mean arterial pressure(MAP),heart rate(HR),and respiratory rate(RR)were recorded at the time of entry(T0),immediately after induction of anesthesia(T1),at the end of gastroscopy(T2),and 5 minutes after awakening(T3).The occurrence of perioperative adverse events and the satisfaction of anesthesiologists and endoscopists were recorded.Results Compared with group C,the incidence of subclinical respiratory depression,hypoxia and severe hypoxia in group M was significantly decreased(P<0.05).The minimum value of SpO2 during anesthesia in group M was higher than that in group C(P<0.05).The incidence of jaw-lifting and mask ventilation were lower in group M than that in group C(P<0.05).The incidence of gastroscope withdrawal due to airway intervention in group M was lower than that in group C(P<0.05).Compared with T0,MAP,HR and RR of the two groups were significant reduction at T1 and T2(P<0.05).There was no significant difference in the incidence of adverse events between the two groups(P>0.05).The satisfaction of endoscopists in group M was higher than that in group C(P<0.05).Conclusion Compared with nasal catheter oxygen inhalation,anesthesia nasal mask can timely detect hypopnea and respiratory depression during gastroscopy,reduce the occurrence of hypoxia,and improve the airway safety of painless gastroscopy patients.

Background With the change of the national energy development layout, Qingyang has seen a situation where oil exploitation and agriculture go hand in hand, which may lead to local soil pollution if not taken seriously. Objective To evaluate the distribution characteristics, possible sources, and ecological risks of heavy metals in farmland soils around the main production areas of Changqing Oilfield. Methods A total of 60 farmland soil samples were collected from Zhengning County, Zhenyuan County, and Qingcheng County of Qingyang City, and the contents of heavy metals such lead (Pb), mercury (Hg), chromium (Cr), cadmium (Cd), and arsenic (As) in farmland soil were detected according to GB 15618-2018 Soil environmental quality—Risk control standard for soil contamination of agricultural land (on trial). The soil background value of Gansu Province was used as the denominator in the calculation of pollution index, and the pollution characteristics and ecological characteristics of selected five heavy metals in farmland soil were evaluated by single-factor pollution index (P_i), Nemerow comprehensive pollution index (P_N), and potential ecological risk index. Results The levels of Pb, As, and Hg in farmland soils around Changqing Oilfield, the levels of Cr and Cd in Qingcheng County, and the level of Cd in Zhengning County were higher than the corresponding soil background values of Gansu Province, but lower than the national soil environmental quality standard. The single-factor pollution indexes (P_i) were: Hg, 2.14; Pb, 1.24; As, 1.13; Cr, 0.78; Cd, 0.67, which indicated that Hg were graded as moderate pollution, Pb and As were slight pollution, and Cd and Cr were not polluted. The Nemerow comprehensive pollution indexes (P_N) were: Cr, 0.92; Cd, 1.08; As, 1.20; Pb, 1.68; Hg, 3.85, which indicated that Cr was graded as no pollution, Cd, Pb and As were mild pollution, and Hg was severe pollution. The variation coefficients of Hg and Cd in Zhenyuan County and that of Hg in Qingcheng County were 60.00%, 50.00%, and 50.00%, respectively, which were all greater than 50%, indicating that the pollution of above heavy metals in the location was subject to human activities. The potential ecological risk indexes (E_r) of Pb, Cr, Cd, As, and Hg were 6.20, 1.55, 20.05, 11.28, and 81.64, respectively, indicating that Hg was graded as strong ecological risk, and the other four heavy metals were mild ecological risk. The comprehensive potential ecological risk index (R_RI) was 124.48. Combined with the potential ecological risk index of Hg, the comprehensive potential ecological risk of the five heavy metals in local farmland soils was considered to be at a strong ecological risk level. Conclusion Although the average values of selected five heavy metals in farmland soils surrounding the main production areas of Changqing Oilfield are qualified with the national soil environmental quality standards, they exceed corresponding soil background values of Gansu Province, and there are signs of human influence and potential ecological risks of different degrees.