Objective To investigate the effect and mechanism of melatonin on the anoikis of melanoma cells.Methods The drug concentration of melatonin inhibiting melanoma cell line B16-F10 was optimized based on the effect on CCK-8 assay.An anti-anoikis of melanoma cell model was developed and divided it into four groups:The blank control group,the TrkB activator group,the melatonin group and the melatonin+TrkB activator group.Calce-in AM/EthD-1 fluorescence double staining was used to detect the anoikis of melanoma cells.Reactive oxygen spe-cies were detected using the fluorescent probe DCFH-DA.Western blot was used to detect the expression of Nrf2 protein and TrkB protein in each group.Results Melatonin significantly inhibited the proliferation of melanoma cells in a time-and dose-dependent manner with IC50 of 1×10-7 μmol/L.Its inhibitory effect was found to be related to in-duction of anoikis of melanoma cells.Melatonin could upregulate the generation of cellular reactive oxygen species(P<0.05),while addition of TrkB activator antagonized this effect.Melatonin could reduce the expression of Nrf2 protein and TrkB protein in melanoma cells(P<0.05),and the addition of TrkB activator could inhibite the effect of melatonin on the expression of Nrf2 protein and TrkB protein(P<0.05).Conclusions Melatonin can inhibit the pro-liferation of melanoma cell line B16-F10 through the mechanism of inducing anoikis.

Objective:To investigate the current use of ulinastatin in the treatment of critically ill children by pediatricians in China.Methods:A anonymous questionnaire survey was conducted among 147 pediatric critical care physicians from 36 hospitals across 16 provinces,autonomous regions,and municipalities in China.The survey content consists of three parts: respondents' basic information, the application status of ulinastatin, and the clinical indicators referenced for evaluating the use of ulinastatin. Descriptive statistical analysis was performed on the collected data.Results:Among the 147 respondents,99.32%(146/147) were from tertiary hospitals；72.11%(106/147) worked in specialized ICUs,and 4.08%(6/147)in emergency medicine departments.A total of 68.03%(100/147) of the physicians reported using ulinastatin in clinical practice.The main diseases for which ulinastatin was used were pancreatitis(26.40%),sepsis and septic shock(23.76%),capillary leak syndrome(21.78%),acute respiratory distress syndrome(8.91%),and disseminated intravascular coagulation(6.27%).A total of 90.00% of physicians combined ulinastatin with other medications,including glucocorticoids(26.82%),albumin(23.51%),plasma(17.22%),and immunoglobulins(13.58%). Clinical indicators referenced during ulinastatin use included elevated interleukin(IL)-6(76.87%),tumor necrosis factor-α(44.22%),IL-8(31.97%),IL-1(19.73%),IL-18(10.20%),blood lactate(59.18%),decreased serum albumin levels(70.07%),increased pleural or peritoneal effusion(67.35%),skin and mucosal edema(65.31%),and elevated thrombomodulin among the four coagulation parameters(58.50%).Conclusion:Ulinastatin is mainly used for the treatment of critical illnesses such as pancreatitis and sepsis.Most physicians combine ulinastatin with other drugs,such as glucocorticoids and albumin.Clinical indicators commonly referenced when using ulinastatin include elevated IL-6,increased lactate,and increased pleural effusion,which suggest a high inflammatory state and endothelial damage.

Objective:To investigate the current use of ulinastatin in the treatment of critically ill children by pediatricians in China.Methods:A anonymous questionnaire survey was conducted among 147 pediatric critical care physicians from 36 hospitals across 16 provinces,autonomous regions,and municipalities in China.The survey content consists of three parts: respondents' basic information, the application status of ulinastatin, and the clinical indicators referenced for evaluating the use of ulinastatin. Descriptive statistical analysis was performed on the collected data.Results:Among the 147 respondents,99.32%(146/147) were from tertiary hospitals；72.11%(106/147) worked in specialized ICUs,and 4.08%(6/147)in emergency medicine departments.A total of 68.03%(100/147) of the physicians reported using ulinastatin in clinical practice.The main diseases for which ulinastatin was used were pancreatitis(26.40%),sepsis and septic shock(23.76%),capillary leak syndrome(21.78%),acute respiratory distress syndrome(8.91%),and disseminated intravascular coagulation(6.27%).A total of 90.00% of physicians combined ulinastatin with other medications,including glucocorticoids(26.82%),albumin(23.51%),plasma(17.22%),and immunoglobulins(13.58%). Clinical indicators referenced during ulinastatin use included elevated interleukin(IL)-6(76.87%),tumor necrosis factor-α(44.22%),IL-8(31.97%),IL-1(19.73%),IL-18(10.20%),blood lactate(59.18%),decreased serum albumin levels(70.07%),increased pleural or peritoneal effusion(67.35%),skin and mucosal edema(65.31%),and elevated thrombomodulin among the four coagulation parameters(58.50%).Conclusion:Ulinastatin is mainly used for the treatment of critical illnesses such as pancreatitis and sepsis.Most physicians combine ulinastatin with other drugs,such as glucocorticoids and albumin.Clinical indicators commonly referenced when using ulinastatin include elevated IL-6,increased lactate,and increased pleural effusion,which suggest a high inflammatory state and endothelial damage.

Objective:To investigate the effect of short-chain fatty acid (SCFA) sodium butyrate (NaB) on the polarization of lipopolysaccharide(LPS) induced M1 type alveolar macrophages and the mechanism of action.Methods:Mouse alveolar macrophages (MH-S) were randomly(random number) divided into control group (Control group), sodium butyrate group (NaB group), LPS group, LPS+NaB group (LB group), and LPS+NaB+adenylate activated protein kinase (AMPK) inhibitor (Compound C) group (LC group).The mRNA expression levels of interleukin 6 (IL-6), interleukin 1β(IL-1β), tumor necrosis factor α(TNF-α), cluster of differentiation 86 (CD86), inducible nitric oxide synthase(iNOS) in MH-S cells, and zonula occludens 1 (ZO-1), tight junction protein 4(Claudin-4), and closed protein(Occludin) in mouse lung epithelial cells (MLE-12) were detected by qRT-PCR;Protein levels of IL-6, IL-1β, and TNF-α in the supernatant of MH-S cell medium were measured by ELISA;Western blot determed the protein expression of AMPK, P-AMPK, nuclear factor E2-related factor 2 (Nrf2), and heme oxygenase 1 (HO-1) in MH-S cells;Expression of M1 type macrophage-associated markers CD86 and iNOS were determined by flow cytometry.Results:(1) qRT-PCR and ELISA results were consistent, M1 type macrophage-associated proinflammatory cytokines IL-6, IL-1β and TNF-α significantly reduced in the LB group after NaB addition compared with the LPS ground (all P <0.05); (2)The results of qRT-PCR and flow cytometry were consistent,compared with the LPS group, the LB group showed a significant decrease in M1 type macrophage-related polarization indicators CD86 and iNOS after NaB addition(all P<0.05); (3) Western blot was used to detect the expression of the AMPK/Nrf2/HO-1 signaling pathway,compared with LPS,the addition of NaB in the LB group enhanced the expression of P-AMPK/AMPK, Nrf2 (nucleus), and HO-1 (all P<0.05); compared with the LB group, the LC group decreased the expression of P-AMPK/AMPK, Nrf2 (nucleus), and HO-1 (all P<0.05);the results of flow cytometry showed that compared with the LPS group, the addition of NaB significantly decreased the expression level of iNOS + in the LB group ( P<0.05); compared with the LB group, the addition of Compound C in the LC group reversed the inhibitory effect of NaB on iNOS + ( P <0.05);(4) The qRT-PCR results of MLE-12 cells showed that compared with the LPS group, the LB group showed a significant increase in Z0-1, Claudin-4, and Occludin after the addition of NaB(all P<0.05). Conclusions:SCFA inhibits LPS-induced polarization of M1-type alveolar macrophages and ameliorates the inflammatory response by activating the AMPK/Nrf2/HO-1 signaling pathway.

Objective     To assess the safety and clinical outcomes of segmentectomy in one- or two-staged video-assisted thoracoscopic surgery (VATS) for bilateral lung cancer. Methods    We retrospectively enrolled 100 patients who underwent VATS segmentectomy for bilateral lung cancer at the Department of Thoracic Surgery of Peking Union Medical College Hospital from December 2013 to May 2022. We divided the patients into two groups: a one-stage group (52 patients), including 17 males and 35 females with a mean age of 55.17±11.09 years, and a two-stage group (48 patients), including 16 males and 32 females with a mean age of 59.88±11.48 years. We analyzed multiple intraoperative variables and postoperative outcomes. Results     All 100 patients successfully completed bilateral VATS, and at least unilateral lung received anatomical segmentectomy. Patients in the one-stage group were younger (P=0.040), had lower rate of comorbidities (P=0.030), were less likely to have a family history of lung cancer (P=0.018), and had a shorter interval between diagnosis and surgery (P=0.000) compared with patients in the two-stage group. Wedge resection on the opposite side was more common in the one-stage group (P=0.000), while lobectomy was more common in the two-stage   group. The time to emerge from anesthesia in the one-stage group was longer than that in the first and second operations of the two-stage group (P=0.000, P=0.002). Duration of surgery and anesthesia were similar between two groups (P>0.05). Total number of lymph node stations for sampling and dissection (P=0.041) and lymph nodes involved (P=0.026) were less in the one-stage group. Intraoperative airway management was similar between two groups (P>0.05). The one-stage group was associated with lower activities of daily living (ADL) scores. Conclusion     Segmentectomy is safe in one- or two-staged VATS for bilateral lung cancer, including contralateral sublobectomy and lobectomy. Duration of surgery and perioperative complications are similar between two groups, but the one-stage group is associated with lower ADL scores. On the basis of comprehensive consideration in psychological factors, physical conditions and personal wishes of patients, one-staged sequential bilateral VATS can be the first choice.

Objective:To evaluate the diagnostic performance of the prostate imaging reporting and data system version 2.1 (PI-RADS v2.1) based on multiparametric MRI (mpMRI) in the detection of clinically significant prostate cancer (csPCa).Methods:A total of 561 patients who underwent prostate mpMRI in the First Affiliated Hospital of Guangxi Medical University from June 2015 to December 2020 due to elevated prostate specific antigen were collected ambispectively. The patients were divided into csPCa group (276 cases) and non-csPCa group (285 cases) according to pathological findings. Prostate were scored according to the PI-RADS v2.1 scoring standard by a junior and a senior radiologist. The prostate volume was measured and the prostate specific antigen density (PSAD) was calculated. The diffusion-weighted imaging and dynamic contrast-enhanced MRI images were processed to measure the quantitative parameters of the index lesion, including apparent diffusion coefficient (ADC), volume transfer constant (K trans) and rate constant (K ep) values. The Mann-Whitney U test was used to compare the difference in parameters between the two groups. The predictors of csPCa were screened by logistic regression analysis. Predictive model of multi-parameter was established. The receiver operator characteristic curves were used to evaluate the efficacy of PI-RADS v2.1 and the model in diagnosing csPCa, and the comparisons of area under the curve (AUC) were conducted by DeLong test. Results:Compared with non-csPCa group, the patients in csPCa group had higher PI-RADS score of senior physician, PSAD, K trans and K ep value, lower ADC value ( Z=-16.69, -12.49, -3.43, -4.67, 13.91, all P<0.001). The PI-RADS scores of senior physician (OR=3.064, 95%CI 2.428-3.866, P<0.001), PSAD (OR=1.554, 95%CI 1.170-2.064, P=0.002) and ADC value (OR=0.095, 95%CI 0.032-0.288, P<0.001) were the predictors of csPCa. The AUC of junior, senior physician PI-RADS and combined prediction model were 0.861 (95%CI 0.830-0.892), 0.895 (95%CI 0.868-0.922) and 0.923 (95%CI 0.898-0.944). The pairwise difference was statistically significant (the PI-RADS score between the junior and senior physicians Z=3.24, P=0.001, the difference between the PI-RADS score of junior physician and prediction model Z=5.54, P<0.001, the difference between the PI-RADS score of senior physician and prediction model Z=4.20, P<0.001). Conclusion:Based on mpMRI, the application of PI-RADS v2.1 by junior and senior radiologists has the high diagnostic efficacy for csPCa, and the multi-parameter model has the best diagnostic efficacy for csPCa.

Objective:To explore the clinical charecteristics, imaging features, therapy and prognosis of stroke in children, and provide help for clinical treatment.Methods:The clinical data of 49 children with stroke were collectedand retrospectively analyzed in the Children′s Hospital of Soochow University from January 1, 2019 to December 31, 2019.Results:A mong the 49 children with stroke, 35 were male and 14 were female, aged 1-178 (65.69 ± 55.22) months; the specific etiologies were cerebrovascular malformation, craniocerebral trauma, tumor, vitamin K deficiencies, infectious diseases, rheumatic immune diseases, hemophilia and congenital heart disease. The first symptoms of stroke were disturbance of consciousness, hemiplegia, convulsions, vomiting and headache. The arterial ischemic stroke (18 cases) were mainly caused by craniocerebral trauma and cerebrovascular malformation. The hemorrhagic stroke (31 cases) were mainly caused by arteriovenous malformation, vitamin K deficiency and tumor. The surgical rate in the arterial stroke group was significantly lower than that in the hemorrhagic stroke group.Conclusions:Traumatic cerebral infarction and intracranial arteriovenous malformation are the main causes of arterial ischemic stroke and hemorrhagic stroke in children. Early diagnosis and treatment can significantly improve prognosis.

Objective:To summarize the effect of thoracoscopy for infants with double aortic arch malformation.Methods:The clinical data of 10 infants with double aortic arch malformation from January 2015 to May 2019 were retrospectively summarized, including 7 boys and 3 girls aged from 2 hours to 8 months, 9 with right arch dominance and 1 with left arch dominance. MRI examination was performed at 35 weeks before birth to understand the relationship between vascular ring and trachea and esophagus. Postnatal surgery was performed when trachea compression exceeded one third, and selective operation was performed around six months after birth if the compression was less than one third.Results:All the 10 cases underwent thoracoscopic surgery, and 1 case underwent thoracotomy because oxygen saturation could not be maintained. The operation time was 80-135 minutes, with an average of 100 minutes. Intraoperative bleeding was about 5-10 ml. Postoperative ventilator support time was 4-14 days. 1 patient was cured and discharged except that he gave up treatment for economic reasons. Postoperative follow-up period was 1-6 months. No difficulty in eating, no difficulty in breathing required tracheal stenosis surgery again. 4 patients needed ventilator support for more than 7 days, 2 patients developed pneumothorax and were cured after drainage. One patient presented hoarseness.Conclusion:Prenatal examination can improve the diagnosis rate of double aortic arch deformity, early intervention follow-up in pediatric surgery can reduce the incidence of related complications in children, and thoracoscopic surgery is feasible to correct the deformity, with small trauma and Less bleeding.

Objective To study the colonization rate and antibiotic resistance of group B Streptococcus (GBS) in gravidas during late pregnancy,and to evaluate the effectiveness of GBS screening in late pregnancy and intrapartum antibiotic prophylaxis (IAP) for the prevention of neonatal early-onset GBS disease (EOGBS).Methods A retrospective study was conducted to analyze the colonization rate and antibiotic resistance pattern of GBS in 14 204 gravidas who were screened for GBS at 35-37 gestational weeks during March 2016 to March 2018 in the University of Hongkong-Shenzhen Hospital (HKU-SZH).Differences in the incidence of EOGBS before and after GBS screening and IAP were analyzed using Chi-square or Fisher's exact test.Results Among the 14 204 gravidas,2 027 cases were GBS positive with a colonization rate of 14.27％.Incidence rates of EOGBS before and after GBS screening were 0.6‰ (4/6 356) and 0.07‰ (1/14 403),respectively (Fisher's exact test,P=0.033).GBS isolates were 100％ (2 027/2 027) sensitive to penicillin and vancomycin.Resistance rates to clindamycin and erythromycin were 67.2％(1 363/2 027) and 65.7％ (1 332/2 027),respectively.Conclusions Routine GBS screening in late pregnancy and IAP can significantly decrease the incidence of EOGBS.Penicillin is the optimal choice for prevention and treatment of GBS infection.

Objective To evaluate the role of nuclear factor kappa B (NF-κB) in cognitive decline in aged mice with sepsis.Methods Forty-five SPF healthy aged female C57BL/6 mice,aged 10-12 months,weighing 20-30 g,were assigned into 3 groups (n=15 each) using a random number table:control group (group C),sepsis group (group Sep) and NF-κB selective inhibitor pyrrolidine dithiocarbamate (PDTC) group (group PDTC).Lipopolysaccharide 250 μg/kg was injected intraperitoneally once a day for 7 consecutive days in Sep and PDTC groups,and in addition PDTC 50 mg/kg was injected intraperitoneally at 30 min before lipopolysaccharide injection once a day for 7 consecutive days in group PDTC.The equal volume of normal saline was given in group C.Five mice in each group were sacrificed at 2 h after the last administration,cardiac puncture was performed and blood samples were collected,and then the mice were sacrificed and hippocampi were harvested for determination of the levels of tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and IL-6 in plasma and hippocampal tissues using enzyme-linked immunosorbent assay.Cognitive function was assessed using open field,elevated plus maze and Morris water maze tests at 24 h after the last administration in the other mice left in each group.Results Compared with group C,the levels of TNF-α,IL-1β and IL-6 in plasma and hippocampal tissues were significantly increased,the time of movement at the central region was shortened,the percentage of time spent in the open arms and number of entries into the open and closed arms were decreased,the escape latency was prolonged,the time of staying at the original platform quadrant was shortened,and the frequency of crossing the platform was decreased in group Sep (P＜0.05).Compared with group Sep,the levels of TNF-α,IL-1β5 and IL-6 in plasma and hippocampal tissues were significantly decreased,the time of movement at the central region was prolonged,the percentage of time spent in the open arms and number of entries into the open and closed arms were increased,the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in group PDTC (P＜0.05).Conclusion NF-κB is involved in cognitive decline in aged mice with sepsis.