Hypoxia Affects Autophagy in Human Umbilical Vein Endothelial Cells via the IRE1 Unfolded Protein Response

Objective:The purpose of this study was to investigate the role of the unfolded protein response,specifically the inositol-requiring enzyme 1(IRE1)signaling pathway,in hypoxia-induced autophagy in human umbilical venous endothelial cells(HUVECs).Methods:The expression of IRE1 and autophagy relative protein in HUVECs with hypoxia was explored by Western blotting,qRT-PCR and confocal microscopy.Further,we evaluated the biological effects of HUVECs by tube formation assay and wound healing assay in vitro.Finally,we examined the function of IRE1 in local blood vessels through animal models,Results:Hypoxia activated the IRE1 signaling pathway and induced autophagy in a time-dependent manner in HUVECs and further influenced the biological effects of HUVECs.Intraperitoneal injection of IRE1 inhibitors inhibited local vascular autophagy levels and lipid accumulation in model animals.Conclusion:Hypoxia can induce autophagy and activate the IRE1 signaling pathway in HUVECs and the IRE1 signaling pathway is involved in autophagy in hypoxic conditions.

Reduced Imaging Time and Improved Image Quality of 3D Isotropic T2-Weighted Magnetic Resonance Imaging with Compressed Sensing for the Female Pelvis

This study is to compare three-dimensional(3D)isotropic T2-weighted magnetic resonance imaging(MRI)with compressed sensing-sampling perfection with application optimized contrast(CS-SPACE)and the conventional image(3D-SPACE)sequence in terms of image quality,estimated signal-to-noise ratio(SNR),relative contrast-to-noise ratio(CNR),and the lesions’conspicuous of the female pelvis.Thirty-six females(age:51,28-73)with cervical carcinoma(n=20),rectal carcinoma(n=7),or uterine fibroid(n=9)were included.Patients underwent magnetic resonance(MR)imaging at a 3T scanner with the sequences of 3D-SPACE,CS-SPACE,and twodimensional(2D)T2-weighted turbo-spin echo(TSE).Quantitative analyses of estimated SNR and relative CNR between tumors and other tissues,image quality,and tissue conspicuity were performed.Two radiologists assessed the difference in diagnostic findings for carcinoma.Quantitative values and qualitative scores were analyzed,respectively.The estimated SNR and the relative CNR of tumor-to-muscle obturator internus,tumor-to-myometrium,and myometrium-to-muscle obturator internus was comparable between 3D-SPACE and CS-SPACE.The overall image quality and the conspicuity of the lesion scores of the CS-SPACE were higher than that of the 3D-SPACE(P<0.01).The CS-SPACE sequence offers shorter scan time,fewer artifacts,and comparable SNR and CNR to conventional 3D-SPACE,and has the potential to improve the performance of T2-weighted images.

Simultaneous determination of doxorubicin and its dipeptide prodrug in mice plasma by HPLC with fluorescence detection

A simple and sensitive high performance liquid chromatography with fluorescence detection(HPLC–FD)has been developed for simultaneous quantification of doxorubicin(DOX) and its dipeptide conjugate prodrug(PDOX) in mice plasma.The chromatographic separation was carried out on an Amethyst C_(18)–H column with gradient mobile phase of 0.1% formic acid and 0.1% formic acid in acetonitrile at a flow rate of 1.0 mL/min.The excitation and emission wavelengths were set at 490 and 550 nm,respectively.The method was comprehensively validated.The limits of detection were low up to 5.0 ng/m L for DOX and 25.0 ng/m L for PDOX.And the limits of quantification were low up to 12.5 ng/m L for DOX and 50 ng/m L for PDOX,which were lower than those for most of the current methods.The calibration curves showed good linearity(R^2>0.999) over the concentration ranges.The extraction recoveries ranged from 84.0% to 88.2% for DOX and from 85.4% to 89.2% for PDOX.Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%.The results show that the developed HPLC–FD method is accurate,reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX.

Effect of fluoxetine on depression-induced changes in the expression of vasoactive intestinal polypeptide and corticotrophin releasing factor in rat duodenum

AIM: To investigate changes in vasoactive intestinal polypeptide (VIP) and corticotrophin releasing factor (CRF) in the plasma and duodenum of chronic stress- induced depressed rats and the effects of fluoxetine hydrochloride (fluoxetine) treatment on depression- induced changes in VIP and CRF. METHODS: A Sprague-Dawley rat model of chronic stress-induced depression was produced. Thirty experimental rats were randomly divided into the following groups: control group, saline-treated depressed group, and fluoxetine-treated depressed group. Open- f ield testing was performed to assess the rats’ behavior. VIP and CRF levels in plasma were measured by ELISA. Immunofluorescence techniques combined with laser scanning confocal microscopy (LSCM) were used to investigate VIP and CRF expression in the duodenum. RESULTS: The open-field behavior, both crossing and rearing, of depression model rats, decreased signif icantly compared with those of normal control rats over 5 min. Defecation times increased significantly. Compared to the control group, FITC fluorescence of duodenal CRF expression and plasma CRF levels in the depressed rats increased significantly (fluorescence intensity of duodenal CRF: 11.82 ± 2.54 vs 25.17 ± 4.63; plasma CRF: 11.82 ± 2.54 ng/L vs 25.17 ± 4.63 ng/L, P < 0.01), whereas duodenal VIP expression and plasma VIP levels decreased signif icantly (fluorescence intensity of duodenal VIP: 67.37 ± 18.90 vs 44.51 ± 16.37; plasmaVIP: 67.37 ± 18.90 ng/L vs 44.51 ± 16.37 ng/L, P < 0.01). Fluoxetine improved depressed behavior, increased VIP expression and decreased CRF expression in plasma and the duodenal tissue of depressed rats. CONCLUSION: Chronic stress can induce injury to the duodenum, accompanied by increasing CRF and decreasing VIP in the plasma and duodenum. Treatment with fluoxetine can ameliorate pathological changes in the duodenum of depressed rats, which suggests that antidepressants are an effective therapeutic agent for some duodenal diseases caused by chronic stress. VIP is a potential therapeutic strategy.

Verification of the Cut-off Value of the Reticulocyte Hemoglobin Content to Diagnose Iron Deficiency

Iron deficiency anemia(IDA)is a common nutrient deficiency,and it has become a global public health problem identified by the World Health Organization.Studies have shown that IDA remains prevalent in both developing and developed countries,such as Australia and China,and that IDA may be associated with poorer cognitive and neurophysiologic maturation in early life and causes labor productivity losses,developmental delays,men tai and behavior disorders,and peri natal complicati ons in pregna nt and preme nopausal women[1].

A retrospective study of patients complaining of nontraumatic acute abdominal pain,admitted in the emergency department of an urban hospital in China

Background:Nontraumatic acute abdominal pain(AAP)accounts for a large proportion of emergency department(ED)admissions.This study aimed to explore the underlying correlations among basic information,imaging examinations,and diagnoses.Methods:A total of 7453 patients complaining of AAP,admitted to the ED of Zhongnan Hospital of Wuhan University,were enrolled in this retrospective study from January 1 to December 30,2019.We collected the following information from the patients:sex,age,date of visit,pain location,nature of pain,level of severity,imaging(computed tomography,x-ray,and ultrasound),diagnosis,and outcomes(re-lease from the hospital,transfer to another hospital,transfer to another department,observation room,hospitalization,or death).Results:According to this study,AAP was more common in female than male.A total of 82.11%patients of AAP were in level 3 of se-verity,while 0.19%patients were in level 1.A total of 77.20%of the patients had undergone imaging.Swelling pain and colic are the 2 most common types of AAP.Nonspecific abdominal pain(NSAP)is the most common diagnosis.In the diagnosis of NSAP and gastroenteritis,female patients were more prevalent than male patients,but for renal colic,male patients were 3 times as many as female patients.Non-specific abdominal pain,biliary colic,and cholecystitis are the 3 leading causes in patients 65 years or older.Nonspecific abdominal pain,renal colic,and gastroenteritis are the 3 leading causes of AAP in patients younger than 65 years.Conclusion:With the help of imaging,clinicians can specify a diagnosis and perform corresponding treatment in most cases.However,making a precise diagnosis of AAP within a short period is still challenging.Further research should be conducted to seek safer and more effective techniques to streamline clinicians’work.

Diagnosis,treatment and prevention of severe acute respiratory syndrome coronavirus 2 infection in children:experts'consensus statement updatedforthe Omicron variant

It has been more than 3 years since the novel coronavirus(SARS-CoV-2)pandemic raged globally.The coronavirus disease 2019(COVID-19)has greatly influenced human society.According to data from the World Health Organization(WHO),there were over 656 million confirmed cases of COVID-19 in the world as of January 1,2023,including over 6.6 million deaths[1].

Diagnosis, treatment, and prevention of 2019 novel coronavirus infection in children: experts' consensus statement

Since the outbreak of 2019 novel coronavirus infection (2019-nCoV) in Wuhan City,China,by January 30,2020,a total of 9692 confirmed cases and 15,238 suspected cases have been reported around 31 provinces or cities in China.Among the confirmed cases,1527 were severe cases,171 had recovered and been discharged at home,and 213 died.And among these cases,a total of 28 children aged from 1 month to 17 years have been reported in China.For standardizing prevention and management of 2019-nCoV infections in children,we called up an experts' committee to formulate this experts' consensus statement.This statement is based on the Novel Coronavirus Infection Pneumonia Diagnosis and Treatment Standards (the fourth edition) (National Health Committee) and other previous diagnosis and treatment strategies for pediatric virus infections.The present consensus statement summarizes current strategies on diagnosis,treatment,and prevention of 2019-nCoV infection in children.

Advances in the Immunotherapeutic Potential of Isocitrate Dehydrogenase Mutations in Glioma

Isocitrate dehydrogenase(IDH)is an essential metabolic enzyme in the tricarboxylic acid cycle(TAC).The high mutation frequency of the IDH gene plays a complicated role in gliomas.In addition to affecting gliomas directly,mutations in IDH can also alter their immune microenvironment and can change immune-cell function in direct and indirect ways.IDH mutations mediate immune-cell infiltration and function by modulating immune-checkpoint gene expression and chemokine secretion.In addition,IDH mutation-derived D2-hydroxyglutarate can be absorbed by surrounding immune cells,also affecting their functioning.In this review,we summarize current knowledge about the effects of IDH mutations as well as other gene mutations on the immune microenvironment of gliomas.We also describe recent preclinical and clinical data related to IDH-mutant inhibitors for the treatment of gliomas.Finally,we discuss different types of immunotherapy and the immunotherapeutic potential of IDH mutations in gliomas.

Detection of an anti-angina therapeutic module in the effective population treated by a multi-target drug Danhong injection:a randomized trial

It's a challenge for detecting the therapeutic targets of a polypharmacological drug from variations in the responsed networks in thedfferentiated populations with complex diseases,as stable coronary heart disease.Here,in an adaptive,31-center,randomized,double-blind trial invoving 920 patients with moderate symptomatic stable angina treated by 14-day Danhong injection(DHI),a kind of polyphamacological drug with high quality control,or placebo(0.9%saline),with 76-day following-up,we firstly confrmed that DHl couldincrease the proportion of patients with clinically significant changes on angina-frequency assessed by Seattle Angina Questionnaire(ASAQ-AF220)(12.78%at Day 30,95%confidence interval[C]5.86-19.71%,P=0.0003,13.82%at Day 6C0,95%CI 6.82-20.82%,P=0.0001and 8.95%at Day 90,95%CI 2.06-15.85%,P=0.01).We also found that there were no significant differences in new-onset major vascularevents(P=0.8502)and serious adverse events(P=0.9105)between DHl and placebo.After performing the RNA sequencing in 62 selectedpatients,we developed a systemic modular approach tp identfy differentilly expressed modules(DEMs)of DHI with the Z_(summay)valueless than 0 compared with the control group,calculated by weighted gene co-expression network analysis(WGCNA),and sketched out thebasic framework on a modular map with 25 functional modules targeted by DII.Furthermore,the effective therapeutic module(ETM),defined as the highest corelation value with the phenotype alteration(SAQ-AF,the change in SAQ-AF at Day 30 from baseline)calculatedby WGCNA,was identifed in the population with the best effect(ASAQ-AF240),which is related to anticoagulation and regulation ofcholesterol metabolism.We assessed the modular flexbility of this ETM using the global topological D value based on Euclidean distance,which is corelated with phenotype alteration(r^(2):0.8204,P=0.019)by linear regression.Our study identified the ant-angina therapeuticmodule in the effective population treated by the multi-target drug.Modular methods facilitate the discovery of network pharmacologicalmechanisms and the advancement of precision medicine.(ClinicalTrials.gov identifier:NCTO1681316).

Can pathologists reliably establish ampulla of Vater carcinoma histologic subtype on H&E alone?Concordance of subtype and comparison to immunohistochemistry-established subtype

Histologic subtyping of ampulla of Vater carcinoma(ampullary carcinoma,AC)may guide choice of adjuvant chemotherapy for advanced disease,as intestinal(INT)subtype may have a better treatment response and survival as compared to the pancreatobiliary(PB)and mixed(MIX)subtypes.However,ambiguous morphologic features can result in inconsistent subtyping among pathologists.Recently immunohistochemistry(IHC)for cytokeratin(CK)7,CK20,CDX2,and the Mucin(MUC)family proteins had been used to aid in the subtyping.It is important to determine if accurate subtyping can be achieved based on hematoxylin and eosin(H&E)evaluation alone,or if IHC is required.In this study,5 gastrointestinal pathologists evaluated 34 cases of AC for histologic subtype(PB,INT,or MIX)based on H&E stain alone.IHC studies(CK7,CK20,and CDX2)were used to determine a gold standard subtyping for comparison(strong CK7 with weak CK20 and CDX2=PB;strong CK20 or CDX2 and weak CK7=INT;strong CK7 and CK20 or CDX2=MIX).Interobserver concordance(kappa)was calculated.Percent correctness of subtyping was calculated in relation to IHC-established subtype gold standard.Interobserver concordance for subtype was fair(kappa 0.31).Percent correctness for subtype against IHC-established subtype was 50%for a majority(3 or more)of pathologists.In all 11 cases with 4 or 5 pathologists being concordant,the subtype was also correctly identified.In addition,data on chemotherapy regimen and overall survival was gathered from the electronic medical record.Patients who were assigned a subtype at time of diagnosis tended to receive gemcitabine for PB subtype,and FOLFOX-based therapy for INT subtype,with some cases diagnosed as"not-otherwise specified"typically also receiving gemcitabine.Survival data were limited by loss to follow-up.In conclusion,despite 11 cases with strong concordance and correctness,overall subtyping concordance was only fair,and a majority of the 5 pathologists correctly identified the subtype in only 50%of cases.Thus,while some AC cases yield clear-cut subtyping,significant discrepancy among pathologists remains.It appears that utilization of IHC to identify subtype in AC cases is necessary to provide accurate,reliable prognostic and therapeutic information.

Pancreatic cystic tumors:an update

Pancreatic cystic tumors(PCTs)comprise a heterogeneous group of entities,accounting for 2%to 10%of pancreatic lesions.The most common types are intraductal papillary mucinous neoplasms(IPMNs),mucinous cystic neoplasm(MCN),and serous cystic neoplasm(SCN),which account for approximately 90%of PCTs.This review discusses updates in pathologic features,malignant transformation,biologic behavior,and molecular evolution of PCTs.IPMN includes main duct and branch duct types.These can also be classified into 4 histologic subtypes based on cell lineages of differentiation,and may be associated with different tumorigenic pathways and clinicopathologic characteristics.The gastric type is the most common and is rarely associated with carcinomas,whereas the pancreatobiliary type is significantly more associated with invasive carcinoma.MCN is a mucinous cystic lesion with the presence of ovarian-type pericystic stroma.Prognosis of the resected non-invasive MCN is excellent,but the long-term survival of MCNs with invasive carcinoma may be poor.SCN includes microcystic adenoma,macrocystic adenoma,and solid variant serous adenoma.Serous cystadenocarcinoma is defined by the presence of distant metastases,which is rare in literature.Intraductal tubulopapillary neoplasm is characterized by uniformly high-grade dysplasia and ductal differentiation without overt production of mucin,with high risk for developing invasion.Acinar cell cystadenoma is a rare benign lesion with acinar differentiation.In addition,some pancreatic neuroendocrine tumors may assume a cystic configuration,sometimes referred to as cystic pancreatic endocrine neoplasm tumor,with a lower pathologic stage.Solid pseudopapillary tumor is composed of poorly cohesive monomorphic epithelial cells forming solid and pseudopapillary structures,with excellent prognosis.

Updated diagnosis, treatment and prevention of COVID-19 in children: experts' consensus statement (condensed version of the second edition)

In the early February,2020,we called up an experts' committee with more than 30 Chinese experts from 11 national medical academic organizations to formulate the first edition of consensus statement on diagnosis,treatment and prevention of coronavirus disease 2019 (COVID-19) in children,which has been published in this journal.With accumulated experiences in the diagnosis and treatment of COVID-19 in children,we have updated the consensus statement and released the second edition recently.The current version in English is a condensed version of the second edition of consensus statement on diagnosis,treatment and prevention of COVID-19 in children.In the current version,diagnosis and treatement criteria have been optimized,and early identification of severe and critical cases is highlighted.The early warning indicators for severe pediatric cases have been summarized which is utmost important for clinical practice.This version of experts consensus will be valuable for better prevention,diagnosis and treatment of COVID-19 in children worldwide.

Expert consensus on COVID-19 vaccination in children

Coronavirus disease 2019(COVID-19)is a public health disaster that has not been encountered for a hundred years.On January 12,2020,the World Health Organization(WHO)confirmed and named the coronavirus which caused unexplained pneumonia as 2019 novel coronavirus(2019-nCoV).Since then,the disease caused by this virus had been named as coronavirus disease 2019(COVID-19).On February 11,2020,the International Committee on Taxonomy of Viruses announced that the English name of the novel coronavirus was severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)[1].At present,SARS-CoV-2 infection is still rampant worldwide.As of September 10,2021,there were about 222 million confirmed cases of COVID-19 and more than 4.5 million deaths worldwide[2].According to the American Centers for Disease Control and Prevention,by July 29,2021,4.19 million pediatric COVID-19 cases had been reported in the United States。