Objective:To evaluate the effect of cathepsin B(CTSB)/NOD-like receptor pyrin domain containing 3(NLRP3)pathway on the polarization of macrophages induced by LPS.Methods:The well-growing RAW264.7 mouse mononuclear macrophage lines were cultured in vitro and divided into 3 groups(n=6)according to the random number table method:control group(C group),LPS group(L group)and LPS+CA074-me(CTSB inhibitors)group(B group).C group was cultured normally for 24 h,L group was cultured with LPS concentration of 1 μg/ml medium for 24 h.B group was pretreated with CTSB inhibitor CA074-me 30 μmol/L for 1 h before LPS induction,and co-cultured with LPS concentration of 1 μg/ml medium for 24 h.After 24 hours,the morphological changes of the cells were observed by microscope,the concentrations of IL-1β and IL-18 in the supernatant were determined by ELISA.The ex-pressions of cathepsin B precursor(pro-CTSB),mature cathepsin B(mature-CTSB),NLRP3,apoptosis-related speck protein(ASC)and apoptosis-related speck protein-1(caspase-1)were detected by Western blot.The mRNA expression levels of CD32,inducible ni-tric oxide synthase(iNOS),arginase 1(Arg-1)and CD206 were detected by qRT-PCR.The positive expression rates of M1 macro-phage surface marker CD86 and M2 macrophage surface marker CD206 were detected by flow cytometry.Results:Compared with group C,the morphology of cells in groups L and B became larger and pseudopodia appeared.The concentrations of IL-1β and IL-18 in cell supernatant were increased,the expressions of pro-CTSB,mature-CTSB,NLRP3,ASC and caspase-1 were increased,and the expressions of CD32,iNOS mRNA were up-regulated and the positive rates of CD86 and CD206 were increased(P<0.01).Arg-1 and CD206 mRNA in group B were up-regulated(P<0.01).Compared with group L,the pseudopodia of group B were reduced,and the morphology was closer to group C.The concentration of IL-1β and IL-18 in the supernatant,the expression of mature-CTSB,NLRP3,ASC and caspase-1,CD32 and iNOS mRNA and the positive rate of CD86 were down-regulated in group B.The expression of pro-CTSB,Arg-1 and CD206 mRNA and the positive rate of CD206 were increased(P<0.01).Conclusion:Inhibition of CTSB/NLRP3 pathway can reduce the inflammatory response,reduce the LPS-induced polarization of RAW264.7 cells to M1 macrophages,and pro-mote their polarization to M2 macrophages.

Objective:To evaluate the effect of cathepsin B(CTSB)/NOD-like receptor pyrin domain containing 3(NLRP3)pathway on the polarization of macrophages induced by LPS.Methods:The well-growing RAW264.7 mouse mononuclear macrophage lines were cultured in vitro and divided into 3 groups(n=6)according to the random number table method:control group(C group),LPS group(L group)and LPS+CA074-me(CTSB inhibitors)group(B group).C group was cultured normally for 24 h,L group was cultured with LPS concentration of 1 μg/ml medium for 24 h.B group was pretreated with CTSB inhibitor CA074-me 30 μmol/L for 1 h before LPS induction,and co-cultured with LPS concentration of 1 μg/ml medium for 24 h.After 24 hours,the morphological changes of the cells were observed by microscope,the concentrations of IL-1β and IL-18 in the supernatant were determined by ELISA.The ex-pressions of cathepsin B precursor(pro-CTSB),mature cathepsin B(mature-CTSB),NLRP3,apoptosis-related speck protein(ASC)and apoptosis-related speck protein-1(caspase-1)were detected by Western blot.The mRNA expression levels of CD32,inducible ni-tric oxide synthase(iNOS),arginase 1(Arg-1)and CD206 were detected by qRT-PCR.The positive expression rates of M1 macro-phage surface marker CD86 and M2 macrophage surface marker CD206 were detected by flow cytometry.Results:Compared with group C,the morphology of cells in groups L and B became larger and pseudopodia appeared.The concentrations of IL-1β and IL-18 in cell supernatant were increased,the expressions of pro-CTSB,mature-CTSB,NLRP3,ASC and caspase-1 were increased,and the expressions of CD32,iNOS mRNA were up-regulated and the positive rates of CD86 and CD206 were increased(P<0.01).Arg-1 and CD206 mRNA in group B were up-regulated(P<0.01).Compared with group L,the pseudopodia of group B were reduced,and the morphology was closer to group C.The concentration of IL-1β and IL-18 in the supernatant,the expression of mature-CTSB,NLRP3,ASC and caspase-1,CD32 and iNOS mRNA and the positive rate of CD86 were down-regulated in group B.The expression of pro-CTSB,Arg-1 and CD206 mRNA and the positive rate of CD206 were increased(P<0.01).Conclusion:Inhibition of CTSB/NLRP3 pathway can reduce the inflammatory response,reduce the LPS-induced polarization of RAW264.7 cells to M1 macrophages,and pro-mote their polarization to M2 macrophages.

Objective:To examined the role of copper starvation in oral squamous cell carcinoma(OSCC),along with the molecular functions and mechanisms of SLC31A1 in these cells.Methods:Initially,SLC31A1 was silenced to induce a copper starvation environment.Sub-sequently,we evaluated the effects of copper starvation on oral squamous cell carcinoma cells using the CCK-8 assay,cell scratch assay,and subcutaneous tumor formation in nude mice.In addition,changes in autophagy and EZH2 expression levels in oral squamous cell carcinoma cells were detected after SLC31A1 silencing.Lactate dehydrogenase activity was assayed to determine its impact on natural killer(NK)cell cytotoxicity after SLC31A1 silencing.Results:Silencing of SLC31A1 significantly inhibited the proliferation,migration,and subcutaneous tu-mor formation ability of oral squamous cell carcinoma cells.Furthermore,silencing SLC31A1 mediated EZH2 degradation and increased NK cell infiltration.Conclusions:Copper starvation can regulate the proliferation,migration,and subcutaneous tumor formation ability of oral squamous cell carcinoma and increase NK cell infiltration by modulating EZH2 expression.

Objective:To investigate the clinical characteristics of family clustering pediatric and adult cases with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant infection in Shanghai City.Methods:A field investigation among the pediatric cases with Omicron variant infection and their household contacts from April 4 to April 30, 2022 in Children′s Hospital of Fudan University was conducted. The informations on case finding, clinical manifestations and SARS-CoV-2 vaccination status were collected. The epidemiological and clinical characteristics were compared between pediatric cases and adult cases. The independent sample t test or chi-square test was used for statistical analysis, and the relative risk ( RR) and 95% confidence interval (95% CI) were used to evaluate the protective effect of vaccination on the infection of Omicron variant. Results:There were 1 274 family members in 297 families including 370 children and 904 adults of whom 1 110(87.13%) were infected with Omicron variant, with 989(89.10%) symptomatic and 121(10.90%) asymptomatic. There were 355 children infected with Omicron variant, of whom 337(94.93%) were symptomatic, and the main manifestations were fever (96.74%(326/337)) and cough (40.36%(136/337)). Only one pediatric case with Rett syndrome developed critically severe pneumonia. A total of 194 pediatric cases had imaging examination, 64(32.99%) showed pulmonary inflammatory lesions. There were 755 adult cases infected with Omicron variant, of whom 652(86.26%) reported symptoms, and the main manifestations were fever (73.16%(477/652)) and cough (49.85%(325/652)). Among symptomatic cases, fever was more common in pediatric cases than in adult cases, while cough was more common in adult cases than in pediatric cases, and the differences were both statistically significant ( χ2=80.87 and 8.04, respectively, both P<0.01). The fever spike was higher in pediatric cases than in adult cases ((39.3±0.7) ℃ vs (38.6±0.6) ℃), and the difference was statistically significant ( t=9.85, P<0.001). The interval from the onset of symptoms to cycle threshold (Ct) value of the nucleic acid of Omicron variant≥35 was longer in pediatric cases than in adult cases ((13.0±3.1) d vs (10.9±3.6) d), and the difference had statistically significance ( t=2.97, P=0.004). Among 160 children aged 3 to 18 years, 54 (33.75%) received two-dose vaccination. Among the 904 adults, 388 (42.92%) received two-dose vaccination and 293 (32.41%) received a booster dose. In the adult cases, the risk of symptomatic infection was reduced by only 8% ( RR=0.92, 95% CI 0.86 to 0.98, P=0.014) following two-dose vaccination, and the risks of fever and cough following booster vaccination were reduced by 42%( RR=0.58, 95% CI 0.49 to 0.67, P=0.001) and 50% ( RR=0.50, 95% CI 0.34 to 0.78, P=0.001), respectively. Conclusions:Secondary attack rate and symptomatic rate of household infection are high in the context of the Omicron variant outbreak in Shanghai. Symptomatic infection is common in children and adults in household setting. Fever is the most common symptom and fever duration is short. Booster vaccination may provide certain protection against common symptoms caused by Omicron variant infection.

Objective:To investigate the serum measles antibody in children with tumor and to provide the clinical evidence for measles vaccination strategy for this special population.Methods:From January 2016 to December 2018, the blood samples of children who were diagnosed with hematological malignancy or solid tumors and received chemotherapy in the Department of Hematology or Oncology Surgery of Children′s Hospital of Fudan University were collected. Enzyme-linked immunosorbent assay was used to quantitatively detect the level of measles IgG antibody, and dynamically monitor the changes of measles antibody level during chemotherapy. Kruskal-Wallis test and chi-square test were used for statistical analysis.Results:A total of 441 children with tumors were enrolled, with the positive rate of measles antibody of 79.1%(349/441), and only 43.3%(191/441) of children had the protective level of IgG antibody. There was a statistically significant difference of the antibody protection rate in children aged0.05). The protection rate of serum measles antibody in children with hematological malignancy and solid tumor were 45.6%(78/171) and 41.9%(113/270), respectively, and there was no statistically significant ( P>0.05). There were 16.3%(16/98) of children who were observed to lose the pre-existing protective antibody during chemotherapy. There was no statistically significant difference of the protection rate of serum among children who had finished chemotherapy 0.05). Conclusions:Serum measles antibody is below the protective level in more than 50% of children with malignancy after chemotherapy. Chemotherapy can compromise the protective antibody against measles. It is recommended for this special population to re-schedule measles vaccine after individualized evaluation to acquire the immuneprotection against measles.

BACKGROUND:Deep vein thrombosis occurred during waiting period before operation affects the prognosis of calcaneal fractures. Therefore, it has important clinical significance for its accurate diagnosis. The staging of thrombosis during waiting period before operation for calcaneal fractures is rarely reported. OBJECTIVE:To detect the incidence and risk factors of deep vein thrombosis in patients with calcaneal fracture by ultrasound elastography imaging after staging diagnosis of thrombosis. METHODS:Al objects were included in the study from patients with calcaneal fractures waiting for surgery in the Qinghai University Affiliated Hospital between 2008 and 2015. Patients received preoperative duplex ultrasonography. Those with thrombosis received ultrasound elastography. The incidence of thrombosis was calculated. According to medical records, age, sex, body mass index and history of smoking were col ected. The correlation between thrombosis and above factors was analyzed by multiple Logistic regression analysis. RESULTS AND CONCLUSION:(1) One hundred and forty-nine patients were final y included. Of these, 22 (14.8%) were found to affect deep vein thrombosis in the lower limb. The incidence of acute thrombosis was 9.4%. (2) Multiple Logistic regression analysis suggested that age (P=0.009, OR=1.063, 95%confidence interval (CI) 1.010–1.117), body mass index (P=0.019, OR=1.302, 95%CI 1.124–1.430), history of smoking (P=0.017, OR=5.124, 95%CI 1.347-18.359), and operation waiting time (P=0.000, OR=5.190, 95%CI 1.396–19.266) were risk factors of acute lower extremity deep venous thrombosis. (3) These results suggest that the incidences of preoperative deep vein thrombosis and acute deep vein thrombosis are very high. The risk assessment of acute deep vein thrombosis should be taken according to the patient’s age, smoking history, body mass index, as wel as the time waiting for surgery. The corresponding diagnosis and treatment program should be given to patients with thrombosis.

Objective To compare the efficacy of balloon dilation and YAG laser endouretertomy in treatment of secondary ureteric stricture . Methods 32 patients with secondary ureteral stricture were randomly divided into balloon dilatation group and holmium laser cut group, 16 cases in each, respectively. The two kinds of treatment efficacy was compared after two years postoperative follow-up. Results The clinical results of the two groups showed no statistical differences at the third, sixth and twelfth month follow-up (P > 0.05), and there were no statistical differences in the overall effective rates (P > 0.05). The clinical results of the two groups showed no statistical difference at the twenty-fourth month follow-up (P < 0.05), and the overall effective rates had statistical difference (P < 0.05). Conclusions Balloon dilation and YAG laser endouretertomy in treatment of secondary ureteral stricture have the same short-term clinical effects. But the middle-term efficacy of YAG laser endouretertomy is superior to that of balloon dilation.

Objective Pulmonary hypertension (PH) was an independent predictor of mortality and new onset cardiovascular events in maintenance hemodialysis (MHD) patients. Recent studies revealed PH complicated by pericardial effusion (PE) increased mortality in non-CKD (chronic kidney disease) population. No such evidence existed in MHD patients. Methods We enrolled 108 MHD patients with diagnosed PH by echocardiography (61 with PE) between 2009 and 2011. All patients had been followed-up for 2 years. The endpoints were all-cause mor-tality, cardiovascular (CV) mortality and new onset CV events. Results The mean age was (60.2 ± 14.1) years, 55.6% were males and SPAP was (53.0 ± 15.4) mmHg. 12.0% were severe PH and 1.9% were moderate-to-severe PE. The overall mortality, cardiovascular mortality and new on-set cardiovascular events in PE and non-PE group were 34.4% vs. 21.3%、 23.0% vs. 12.8% and 54.1% vs. 42.6%, respectively (P > 0.05). Kaplan-Meier curve showed there was no difference on all-cause mortality , CV mortality nor new-onset CV events between PE and non-PE group (Log Rank P > 0.05). Conclusions Compared with patients without PE, the presence of PE in patients with PH did not increase the mortality and worsen the cardiovascular outcome in MHD patients.

Objective To analyze the complications of total proctocolectomy with ileal pouch-anal anastomosis (TPC-IPAA) in treatment of severe or refractory ulcerative colitis (UC).Methods The retrospective cross-sectional study was conducted.The clinical data of 67 patients with severe or refractory UC who underwent TPC-IPAA in the Xinhua Hospital Affiliated to Shanghai Jiaotong University from February 2008 to October 2015 were collected.All the patients received open and laparoscopy-assisted TPC-IPAA,and two-stage or three-stage surgery was performed according to the patients' conditions.Observation indicators:(1) treatment and follow-up situations,(2) short-term complications after TPC-IPAA,(3) long-term complications after TPC-IPAA.The follow-up using telephone interview and outpatient examination was performed to detect the quality of postoperative life in patients and occurrence of complications up to July 2016.Measurement data with skewed distribution were described as M (range).The comparison of quality of pre-and post-operative life in patients was done using the paired t test.Results (1) Treatment and follow-up situations:all the 67 patients received successful surgery,including 45 with two-stage surgery and 22 with three-stage surgery.Thirty-seven patients underwent open surgery and 30 underwent laparoscopic surgery.The average frequency of postoperative daily defecation,average scores of quality of pre-and post-operative lifes were 5.6,0.47 and 0.67,respectively,with a statistically significant difference between preoperative indicators and postoperative indicators (t =-4.80,P ＜ 0.05).All the 67 patients were followed up for a median time of 4.6 years (range,1.0-8.4 years).(2) Short-term complications after TPC-IPAA:Of 67 patients,10 had short-term complications (some patients with multiple complications).One patient was complicated with presacral abscess secondary to sinus formation and then was cured by topical incision of sinus under colonoscopy.Five patients were complicated with anastomotic site-related complications of TPCIPAA,and 1 with pouch-anal anastomotic fistula combined with presacral abscess underwent drainage with ileostomy and didn't undergo stoma reversion of ileum.Two patients with fistula at the top of pouch were improved by surgery and repair.Of 2 patients with pouch-vagina fistula,1 underwent resection of pouch stump-vagina fistula and then were cured,and the space between pouch stump and posterior fornix was filled with a free greater omentum flap and temporary ileal bypass was simultaneously conducted.The other patient received twice transanal vaginal fistula repairs with advancement flap,and unclosed fistula was confirmed by angiography,with a clinical symptomatic relief.Eight patients with wound infection received actively dressing change after discharge,and no wound dehiscence was occurred.(3) Long-term complications after TPC-IPAA:28 of 67 patients had long-term complications.Twelve patients with intestinal obstruction underwent conservative treatments of gastrointestinal decompression,anti-inflammatory with corticosteroid and anti-infection with antibiotic,without the occurrence of severe intestinal perforation and prognosis needing surgical intervention,including 9 undergoing open TPC-IPAA and 3 undergoing laparoscopic TPC-IPAA.Thirteen patients with postoperative pouchitis received the conservative treatment of mesalazine.Three patients with failed pouch underwent small intestine permanent colostomy,including 2 with secondary pouch Crohn's disease and 1 with severe pouchitis.Conclusions TPC-IPAA is safe and effective in treatment of UC,with a lower incidence of complications.Anastomotic site-related complication of TPC-IPAA is the main short-term complication.And in the long-term complications,pouchitis is the most,followed it is intestine obstruction.

Objective To analyze the status of biobanks in public hospitals in Shanghai.Methods A questionnaire survey on biobanks was conducted in 9 representative public hospitals in Shanghai.Results The management system of hospital biobanks in Shanghai was basically shaped,but the human resources and financial inputs were in shortage,and some management and regulations were not in place yet.Conclusions The biobanks of public hospitals need more inputs,improvement,and standardized management.