Clinicopathological analysis of primary nasopharyngeal melanomas in elderly patients / 中华老年医学杂志

Objective:To investigate the clinicopathologic features and molecular genetics characteristics of sinonasal tract mucosal malignant melanomas(STMMMs)in elderly patients.Methods:The clinicopathological features, immunohistochemical features and BRAF, C-KIT, NRAS mutations of STMMM in ten elderly patients were retrospectively analyzed.Results:Among the 10 patients, 5 were female and 5 were male.The patients were aged 65-81 years, with an average age of(72.5 ± 8.5)years.The lesions in 7 cases were located in the nasal cavity and paranasal sinuses, and in the other 3 cases were located in the nasopharynx.The morphologies of tumor cells under microscope was complex and diverse, showing plasma cell-like, rhabdomyoblast-like, small cell-like, epithelial-like, and spindle cell-like morphologies.Immunohistochemically, HMB-45 and S-100 were generally positive in 10 cases, and the positive rate of Melan A was 70.0%.The genes detection data showed no mutations in BRAF or NRAS genes in all the 10 cases, while C-KIT exon 11 c. 1666_1667insA mutation was found in one case, and the remaining 9 cases were wild-type for C-KIT.All the 10 cases were followed up for 4~50 months.Three cases survived so far.Conclusions:STMMM in elderly patients are rare and easy to be misdiagnosed.Immunohistochemistry and genetic testing provide guidance for accurate diagnosis and targeted therapy.

Research progress on the relationship between COVID-19 and autoimmune diseases / 中华预防医学杂志

Different autoantibodies can be detected in patients with coronavirus disease 2019 (COVID-19). It is reported that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection could induce autoimmune diseases (AID), including children's multisystem inflammatory syndrome (MIS-C), Guillain Barre syndrome (GBS), Autoimmune hemolytic anemia (AIHA), immune thrombocytopenia (ITP) and thyroid autoimmune diseases. This article mainly reviews the similarities between COVID-19 and AID, the possibility of COVID-19 inducing AID, the risk of AID patients infected or vaccinated against COVID-19. The purpose is to provide strategies for the prevention, management and treatment of AID during the epidemic.

Clinicopathological features of adenosquamous carcinoma of the lung in elderly patients / 中华老年医学杂志

Objective:To investigate the clinicopathological features, immunohistochemical phenotypes and molecular characteristics of adenosquamous carcinoma of the lung(ASC)in elderly patients.Methods:Clinical data of 72 ASC patients in the Department of Pathology, The Second Affiliated Hospital of Soochow University from January 2009 to December 2020 were retrospectively analyzed, and 48 patients aged ≥60 years were selected.Clinical manifestations, imaging findings, histopathological and immunohistochemical characteristics were collected, and gene mutations were detected by the amplification refractory mutation system(ARMS-PCR).Results:There were 48 patients including 32 males and 16 females with a mean age of 70 years(range: 60-84 years). The maximum diameters of the tumors ranged from 0.3 to 9.0 cm(mean: 2.8 cm). Microscopically, the tumors contained two components, squamous cell carcinoma and adenocarcinoma, with the squamous cell carcinoma tissue showing intercellular bridges and the adenocarcinoma tissue showing papillary, acinar or tubular structures.Immunohistochemistry assays detected varying expression levels of CK7(30/31), CK5/6(20/28), TTF1(12/31), P40(15/17), and P63(12/13). Molecular testing showed that the EGFR mutation rate was 58.8%(10/17)and the ALK fusion mutation rate was 5.9%(1/17), while ROS1 and MET mutations were not detected.All 48 patients underwent surgical resection.Conclusions:ASC cases are relatively rare and prone to misdiagnosis.The diagnosis requires the combination of HE morphology, immunohistochemistry and imaging examination, and surgery is the main treatment option.The mutation rate of the EGFR gene is relatively high in ASC patients.

The correlation between the high expression of C/EBPγ and the proliferation and migration of liver cancer cells / 中华检验医学杂志

Objective:To investigate the levels of C/EBPγ mRNA in liver cancer patients and the effect and mechanism of C/EBPγ on proliferation and migration of HepG2 cells.Methods:This experiment was performed in Department of Clinical Laboratory of Shanghai Songjiang District Central Hospital in vitro from October of 2018 to April of 2019. The pLKO.1 was the control group and shg1/shg2 were interference groups. The transcriptional levels of C/EBPγ in liver cancer were analyzed through Oncomine database and real-time PCR. HepG2 and Hep3B cells were cultivated in DMEM media with 10% FBS at 37℃ and 5% CO 2. Adenovirus vector shC/EBPγ-pLKO.1 was constructed and infected 293T cell combined with packaging carrier pMD2G, pMDLG/REE and pRSV/Rev. The empty vector pLKO.1 served as control. The virus supernatant was collected at 12 h and infected HepG2 cells four times. Puromycin were added at the concentration of 2 μg/mL and cells were selectively cultured for 48-72 h. The puromycin concentration was changed into 1 μg/mL after the selection. Growth curves were used to detect cell proliferation. ROS levels were determined by using DFH-DA assays. The oxidative stress related genes transcription changes were detected by using real-time PCR. The migration of HepG2 cells was assayed through scratch assay. NAC (1 mmol/L) were added to cells against oxidative stress. Results:The mRNA levels of C/EBPγ were much higher in liver cancer patients and cells than normal controls. The proliferation rates of C/EBPγ interference groups at 4 d (shg1: 1.93±0.70, shg2: 1.28±0.40) and 6 d (shg1: 3.05±0.90, shg2: 1.31±0.60) were lower than control groups (4 d: 6.18±0.80, 6 d: 22.41±2.56) significantly ( F=1 507.971, P<0.05) . Reactive oxygen species (ROS) levels were elevated in C/EBPγ interference groups and the transcription level of oxidative stress related genes (HPRT1, NQO1-tv2 and NQO1-tv4) was increased at 12 h and 24 h when the cells were cultured without serum. The scratch assays showed that non-healing area in C/EBPγ interference groups (shg1: 174 922.58±1 239 376.00,shg2: 1 374 656.00±248 882.79) was larger than control groups (66 690.82±1 278 954.00) ( F=39.871, P<0.05) . When the antioxidant NAP was added, the proliferation rates were elevated at 6 h and the ROS levels were decreased at 0.5 h, 1 h and 3 h ( F=7.587, 4.657, 3.903, P<0.05) . Conclusions:C/EBPγ mRNA levels were increased in liver cancer. When C/EBPγ was disturbed in HepG2 cells, the proliferation and migration were decreased, the ROS levels and oxidative stress related genes were elevated significantly, which indicated that C/EBPγ promoted proliferation and migration through reducing ROS levels in liver cancer.

Hepatitis B virus x protein promote human hepatocellular carcinoma cells proliferation by activating p38 and JNK signalling pathway / 中华实验和临床病毒学杂志

Objective@#To investigate the effects and mechanism of hepatitis B virus x protein (HBx) on human hepatocellular carcinoma cells proliferation.@*Methods@#Eukaryotic expression vector HBx-pEGFP-C1 was constructed. HepG2 cells were transfected transiently using Lipofectamine 2000. HBx expression in transfected cells were measured by RT-PCR and Western blot. Cells proliferation and apoptosis were detected by using growth curves and TUNEL staining. The protein levels of caspase-3, p-p38, p-Akt and p-JNK were measured by Western blot.@*Results@#HBx was successfully expressed in HepG2 cells. Growth curve result showed that HBx promoted cell proliferation (t=-0.8999, P=0.012). Compared with control group, the levels of p-p38(24 h) (t=- 11.058, P=0.0004) and p-JNK(48 h) (t=- 15.022, P=0.0001) in HBx-pEGFP-C1 group were increased significantly. There is no significant difference between the two groups′ apoptosis.@*Conclusions@#Transient overexpression of HBx promoted human hepatic carcinoma cells proliferation and activated the p38 and JNK signalling pathway.

Advances in CncRNAs research / 中华检验医学杂志

RNAs are known to regulate diverse biological processes, either as protein-encoding molecules or as non-coding RNAs.However, a new class of bi-functional RNAs, carrying both protein coding and RNA-intrinsic functions, have been identified and termed as 'CncRNAs'.This review discuss three major genomic sources of CncRNAs and describe the dual characteristics and functional mechanisms of them, providing a new perspective to further understand the complexity of the transcriptomics and genomes and the complex gene-regulatory network in organisms.

Effects of pioglitazone pre-treating on acinar cells apoptosis induced by caerulein in acute pancreatitis / 中华消化杂志

Objective To investigate the effects of pioglitazone pre-treating on pancreatic acinar cell (AR42J cells) apoptosis induced by caerulein.Methods AR42J cells were divided into blank control group (with normal culture),pioglitazone group (40 μmol/L),caerulein control group (1 × 10-8 mol/L),pioglitazone+ caerulein group (40 μmol/L pioglitazone + 1 × 10-8 mol/L caerulein) and pioglitazone + GW9662+caerulein group (40 μmol/L pioglitazone+ 5 μmol/L GW9662 + 1 × 10-8 mol/L caerulein).Pioglitazone and GW9662 were added 30 minutes earlier than caerulein.Cell proliferation rate of each group was determined by MTT assay at three,six,12 and 24 hour.The cell apoptosis rate was detected by flow cytometry with Annexin Ⅴ/PI staining and terminal dexynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) staining.The activity of Caspase 3,8 and 9 of each group was measured.Mitochondrial membrane potential (MMP) was detected by flow cytometry with JC-1 staining.Single factor analysis of variance and LSD test were performed for data analysis.Results At six,12 and 24 hour,the cell proliferation rate of pioglitazone group and pioglitazone + caerulein group was 0.19±0.02,0.22±0.02,0.36±0.02 and 0.20±0.04,0.23±0.02,0.38±0.02,respectively,which were significantly lower than those of blank control group (0.25 ±0.04,0.28 ± 0.03 and 0.46±0.02) and caerulein group (0.23±0.02,0.29±0.01 and 0.46±0.05,t lgroup=-3.16,-4.61 and-6.25,tcaerulein group =-1.58,-4.61 and-6.15,all P＜0.05).And the cell proliferation rates of pioglitazone+GW9662+caerulein group at six,12 and 24 hour (0.23±0.02,0.27±0.02 and 0.45±0.01) were significantly higher than those of pioglitazone+caerulein group (t=2.25、3.87、4.56,all P＜0.05).There was no significant difference in cell apoptosis rate detected by flow cytometry with Annexin Ⅴ/PI staining between pioglitazone group ((11.80 ± 0.47) ％,(9.62 ± 2.63) ％ and (14.92 ± 2.52) ％) and pioglitazone+caerulein group ((8.78±0.47)％,(11.89±2.80)％ and (14.25±2.67)％,all P＞0.05),but cell apoptosis of these two groups were higher than those of control group ((5.52± 0.64)％,(5.30±0.97)％ and (5.47±0.88)％) and caerulein group ((5.98±1.21)％,(7.47± 0.58) ％ and (8.11 ± 1.32) ％) respectively,and the differences were statistically significant (t l group =9.81,4.45 and 10.74,tcaerulein group =4.38,7.62 and 6.98,all P ＜0.05).There was no significant difference in apoptosis rate between pioglitazone+GW9662+caerulein group ((5.82±0.26) ％,(6.05± 0.83) ％ and (9.23±0.90)％) and caerulein group; while significantly higher when compared with those of pioglitazone+ caerulein group (t=-4.63,-10.07 and-5.70,all P＜0.05).At 12 hour,the apoptosis rate detected by TUNEL staining of pioglitazone group ((3.93 ± 0.40)％) was significantly higher than that of control group ((2.73 ±0.68) ％),the apoptosis rate of pioglitazone+ caerulein group ((8.43 ± 1.65)％) was significantly higher than that of caerulein group ((2.80 ± 0.56)％),the apoptosis rate of pioglitazone+GW9662+caerulein group ((3.87±0.35)％) was lower than that of pioglitazone+ caerulein group (t=7.93,8.92,-5.35,all P＜0.05).At 24 hour,the activity of Caspase 3,8 and 9 of pioglitazone+ caerulein group (1.28 ± 0.05,1.38 ± 0.04 and 1.53 ± 0.09) significantly increased compared with those of caerulein group (1.12±0.88,1.22±0.02 and 0.53±0.07,t=3.20,8.62 and 1.29,all P＜0.05).After treated with GW9662,part of activity of Caspase enzymes recovered.The number of cells with potential change of mitochondrial membrane in pioglitazone group and pioglitazone + caerulein group was more than that of caerulein group (28.50±0.91)％ and (28.20±2.56)％ vs (15.00±3.67)％) and part of membrane potential recovered after GW9662 added ((20.67 ± 2.20) ％).Conclusions Pioglitazone might promote AR42J cell apoptosis through the activation of caspases enzymes and changing membrane potential.And the antagonist GW9662 would partially inhibit the apoptosis induced by pioglitazone.

Mechanism on the role of caspase recruitment domain-containing protein 9 in the early stage of acute pancreatitis / 中华消化杂志

Objective To investigate the mechanism of caspase recruitment domain‐containing protein 9 (CARD9) in the early stage of acute pancreatitis(AP) .Methods Peripheral blood mononuclear cells (PBMC ) of 49 AP patients (33 mild acute pancreatitis (MAP ) patients and 16 severe acute pancreatitis (SAP) patients) were collected on the Day 1st ,3rd and 5th of hospitalization .Twenty healthy volunteers were enrolled in control group .The expression level of CARD9 ,B‐cell lymphoma(Bcl)‐10 ,p38 mitogen‐activated protein kinase (MAPK ) and p65 nuclear factor Kappa B (NF‐κB ) in PBMC of AP patients were detected by Western blotting .The co‐localization ,expression and binding between CARD9 and Bcl‐10 in PBMC of control group ,SAP group and MAP group on the Day 1st hospitalization were determined by cell immune‐fluorescence staining and co‐immuno precipitation method .Single factor analysis of variance and Mann‐Whitney test were performed for data comparison between groups .Pearson method was used for correlation analysis .Results The results of Western blotting indicated that the expression of CARD9 and Bcl‐10 in PBMC of SAP group on the Day 1st ,3rd and 5th of hospitalization (1 .12 ± 0 .05 ,1 .03 ± 0 .03 and 1 .01 ± 0 .01 ;1 .74 ± 0 .08 ,1 .72 ± 0 .10 and 1 .69 ± 0 .11) were all significantly higher than those of control group (0 .33 ± 0 .10 and 1 .02 ± 0 .11) and MAP group (0 .71 ± 0 .02 ,0 .55 ± 0 .06 and 0 .25 ± 0 .07 ;1 .15 ± 0 .03 ,1 .09 ± 0 .07 and 1 .01 ± 0 .04) ,and the differences were statistically significant (F= 35 .76 and 18 .20 ,all P< 0 .05) .The expression of p38 MAPK in PBMC of SAP group on the Day lst ,3rd of hospitalization (1 .88 ± 0 .08 、1 .68 ± 0 .11) were significantly higher than those of MAP group on the Day 1st ,3rd ,5th (0 .86 ± 0 .08 ,0 .77 ± 0 .10 ,0 .73 ± 0 .20) and healthy control group (0 .58 ± 0 .24 , F= 7 .24 ,all P < 0 .01) .The expression of p65 NF‐κB in PBMC of SAP group on the Day 1st ,3rd of hospitalization (1 .64 ± 0 .02 ,1 .55 ± 0 .03) were significantly higher than those of MAP group on the Day 3rd ,5th (1 .06 ± 0 .14 ,0 .87 ± 0 .20) and healthy control group (1 .17 ± 0 .13 ,F= 4 .51 ,all P< 0 .05) .The results of immune‐fluorescence staining indicated that CARD9 and Bcl‐10 co‐localized in nucleus .The results of co‐immuno precipitation showed that the binding degree between CARD9 and Bcl‐10 of SAP group was significantly higher than that of control group and MAP group . Pearson correlative analysis suggested that the level of p65 NF‐κB and p38 MAPK in PBMC of AP patients were positive correlated with the expression of CARD9 (r= 0 .692 and 0 .834 ,both P< 0 .01) .Conclusion CARD9 is positive correlated with NF‐κB and MAPK , which indicates CARD9 induced inflammatory cytokines by activating NF‐κB and MAPK signaling pathways in AP .

A preliminary analysis of plasma microRNA expression profile in patients with primary biliary cirrhosis / 中华检验医学杂志

Objective To evaluate the expression and clinical significance of miRNA in plasma of patients with primary biliary cirrhosis.Methods Plasma from 19 PBC patients,10 healthy volunteers and 10 viral hepatitis patients were selected from Shanghai Songjiang Hospital from december 2010 to January 2013.Among them 3 PBC patients' plasma and 3 healthy volunteers' plasma were detected by miRNA microarray for miRNA expression profile examination.Real-time PCR was used to verify the results of microarray,miRNA target gene predictior software was used to predict the target genes of differentially expressed miRNA.ROC was used to determine the clinical value of plasma miRNA.Results According to microarray,a total of 16 miRNAs were found to be differentially expressed.As revealed by qRT-PCR,the expression of miRNA-92a-3p and miRNA-4516 decreased while the expression of miRNA-572 and miRNA-575 were up-regulated in PBC group compared with healthy controls (P ＜ 0.05).In comparison with nonPBC cirrhosis group,only miRNA-92a-3p and miRNA-4516 were down-regulated (P ＜ 0.05).The area under the curve (AUC)of miRNA-92a-3p for the diagnosis and differential diagnosis of PBC were 0.92 and 0.84,respectively.while for The area under the ROC curve of miRNA-4516,the AUC for diagnosis and differential diagnosis PBC were 0.89 and 0.76,respectively.The optimal cut-off values for identifying PBC from healthy controls were defined as 1.26 ng/μl.for miRNA-92a-3p (sensitivity,92％ ;specificity,80％)and 1.16ng/ul for miRNA-4516 (sensitivity,85％ ;specificity,70％)respectively.The optimal cut-off values for identifying PBC from viral hepatitis were defined as 1.08 ng/μl.for miRNA-92a-3p (sensitivity,89％ ; specificity,81％)and 1.06 ng/μl for miRNA-4516 (sensitivity,77％ ;specificity,68％).Conclusion The results indicate that plasma from patients with PBC has a unique miRNA exprssion profile and these differentially expressed miRNA can be used as clinical biomarkers of PBC.

Study of the serum quality controlling method / 国际检验医学杂志

Objective To improve the serum quality controlling level ,in order to regularize and standardize the laboratory processes .Methods Gallery of hemolysis ,lipidemia or icterus serum specimens was established by Roche cobas p612 QSI camera system .The trouble serum specimens were screened out through taking photos of interfere serum and comparing with the gallery . The serum indexes of hemolysis ,lipidemia and icterus were detected respectively as reference information of inspection report .Com‐paring the results of 16 710 specimens detected by system screening and visual judgment ,the accuracy of QSI camera system was verified .Results 195 trouble serum pictures were elected in the gallery .The compliance rates of hemolysis ,lipidemia and icterus se‐rum specimen between QSI camera system and visual judgment were 94 .2% ,87 .4% ,and 60 .9% respectively .Conclusion The QSI camera system can replace visual judgment to screen the trouble serum specimens .

Effect of pioglitazone on pancreatic apoptosis in rats with acute necrotizing pancreatitis / 中华胰腺病杂志

Objective To investigate the effect of pioglitazone on the activation of pancreatic apoptosis in the pathogenesis of rats with acute necrotizing pancreatitis.Methods Eighty Sprague-Dawley (SD) rats were randomly divided into four groups,including acute necrotizing pancreatitis (ANP),sham operation (SO),solvent control (Solvent),pioglitazone intervention (pioglitazone) group,with 20 rats in each group.ANP model was induced by retrograde injection of 4％ sodium taurocholate (1ml/kg body weight) into the biliary-pancreatic duct.The rats in pioglitazone group were injected pioglitazone (40 mg/kg body weight) into the ANP abdominalcavity 30 min before mldel induction.The rats were sacrificed at 1 h,3 h,6 h,and 12 h after ANP model induction.The pancreatic tissues were harvested.Routine HE staining was used to evaluate pancreatic pathological damage.The apoptosis was determined by TUNEL method.The expression of PPARγ was determined by using immunohistochemistry and Western-blot methods.The activity of caspase3 in pancreatic tissues was detected by using spectrophotometry.Results The pancreatic pathological damage was attenuated in rats in pioglitazone group compared with that in rats of ANP group,and the difference between the two groups was statistically significant (P ＜ 0.05).The PPARγ expression of pioglitazone group was 1.34 ± 0.09,which was significantly higher than that in ANP group (0.75 ± 0.05),and the difference between the two groups was statistically significant (P ＜ 0.05).The apoptotic index in pioglitazone group at 3 h was 8.35 ± 0.95,which was significantly higher than that in ANP group at 3 h (4.37 ± 1.22) ; the caspase3 activity was 9.24 ± 1.78,which was significantly higher than that in ANP group (5.04 ± 0.86),and the difference between the two groups was statistically significant (P ＜0.05).Conclusions Pioglitazone intervention attenuates pancreatic inflammation,increases PPARγ expression and caspase3 activity and induces apoptosis in pancreas of rats with acute necrotizing pancreatitis.

Mechanisms of phagocytosis of Leptospira by peritoneal macrophages of guinea pigs / 上海交通大学学报（医学版）

Objective To investigate the mechanisms of phagocytosis of virulent Leptospira by peritoneal macrophages of guinea pigs,andevaluatetheroleof innateimmuneinthepathogenesisof leptospirosis. Methods Peritoneal macrophages of guinea pigs were extracted. Three specific inhibitors ( microfilament inhibitor cytochalasin D,microtube inhibitor colchicine and PI3K signalling pathway inhibitor LY294002) were added respectively to the macrophages 1 h before the infection of virulent Leptospira interrogans serovar Lai type strain Lai in vitro.Meanwhile, control group without inhibitor was established.Phagocytosis was observed by laser scanning confocal microscopy and phagocytic rates were evaluated by flow cytometry 3 h after infection.ResultsThe phagocytic rates of control group, cytochalasin D group, colchicine group and LY294002 group were (38.98 ± 0.91)%,(23. 99 ± 1. 40) % ,(40.81±0.91)% and (39.64 ±3.56) %, respectively.The phagocytic rate of cytochalasin D group was significantly lower than that of control group (P < 0. 05), while those of colchicine group and LY294002 group were not significantly different from that of control group (P >0.05). ConclusionMicrofilaments play an important role in the phagocytosis of strain Lai by peritoneal macrophages,but the process is independent on PI3K signalling pathway,and microtubes play little part during the phagocytosis.

Cloning of rat serum amyloid P component gene and preparation of its recombinant adenovirus vector / 重庆医科大学学报

Objective:To construct the recombinant adenovirus vector carrying rat serum amyloid P component(SAP)as preparation for later use.Methods:Rat SAP was obtained by using RT-PCR amplification and then cloned into the shutter plasmid pAdTrace-TO4.Subsequently,this newly constructed plasmid pAdTrace-TO4-SAP was followed by homologous recombination with bone plasmid pAdEasy-1 in BJ5183.Recombinant adenovirus was obtained after being packaged in human embryonic kidney cells HEK293 by lipofectAmineTM2000 mediation.After amplification the recombined adenovirus,we determined its titer by end-point dilution assay.Results:In recombinant shuttle plasmid and recombinant adenovirus,a fragment of 700 bp detected by PCR was identical with that included in Genbank.The titer of recombinant adenovirus reached 2.8?108 pfu/ml.Conclusion:The recombinant adenovirus containing SAP gene was constructed successfully.This will provides a good basis for further study about the influence of SAP on lung fibrosis.