The corona virus disease 2019(COVID-19)infection has assumed global pandemic status in a short period of time since its outbreak.This public health crisis has inflicted unprecedented damage and challenges to more than 200 countries and regions around the word.As the peak of the epidemic gradually subsides,an increasing amount of attention has been shifted to the issue of long-term sequelae in COVID-19 patients.These sequelae are manifested as the continuous dysfunction of multiple organ systems and are known as Long COVID.In response to this new issue,clinical pathological research has continuously made new progress,posing new requirements and challenges for the pre-vention and treatment of Long COVID.This article aims to review the progress of clinical pathological research on Long COVID in recent years,with the expectation of providing new ideas and methods for the prevention and treatment of this disease.

The corona virus disease 2019(COVID-19)infection has assumed global pandemic status in a short period of time since its outbreak.This public health crisis has inflicted unprecedented damage and challenges to more than 200 countries and regions around the word.As the peak of the epidemic gradually subsides,an increasing amount of attention has been shifted to the issue of long-term sequelae in COVID-19 patients.These sequelae are manifested as the continuous dysfunction of multiple organ systems and are known as Long COVID.In response to this new issue,clinical pathological research has continuously made new progress,posing new requirements and challenges for the pre-vention and treatment of Long COVID.This article aims to review the progress of clinical pathological research on Long COVID in recent years,with the expectation of providing new ideas and methods for the prevention and treatment of this disease.

Objective To investigate the effect of glycine N-methyl transferase (GNMT)on intrauterine adhesion (IUA)fibrosis and its related mechanism.Methods In vivo experiment:A total of 36 healthy female SD rats (SPF grade,6～8 weeks old and weighing from 180～220 g)were subjected in this study.IUA model of SD rats and IUA model of GNMT overexpressed rats were established.RT-qPCR and immunofluorescence assay were applied to detect GNMT expression level in normal uterus and model group.RT-qPCR and Western blotting were used to detect the mRNA and protein levels of fibrosis-related molecules and the activation of TGF-β1/Smad3 signaling pathway in each group.The number of endometrial glands in each group was observed by HE staining.Masson staining was used to analyze the severity of endometrial fibrosis in each group.In vitro experiment:transformed human endometrial stromal cells (THESCs)fibrotic phenotype model was constructed using TGF-β1,and THESCs stably transfected with GNMT overexpression lentvirus were treated with TGF-β1.RT-qPCR and Western blotting were used to detect the mRNA and protein expression of fibrosis-related molecules.The expression of TGF-β1/Smad3 signaling pathway was detected by Western blotting.TGF-β1/Smad3 signaling pathway was activated by TGF-β1/Smad signaling pathway activator (SRI-011381),and the expression of TGF-β1/Smad3 signaling pathway and key molecular proteins of fibrosis phenotype was measured with Western blotting.Results In vivo experiment,the mRNA and protein expression levels of GNMT were significantly decreased in the IUA rats than the control rats (P<0.05).Overexpression of GNMT decreased the mRNA and protein levels of fibrosis related molecules,Collagen Ⅰ,Collagen Ⅲ and FN in the IUA rats (P<0.05),and decreased the phosphorylation levels of TGF-β1 and its downstream Smad3 protein (P<0.05).HE and Masson staining showed that overexpression of GNMT could increase the number of endometrial glands and reduce the severity of fibrosis in the IUA rats (P<0.05).In vitro experiments:overexpression of GNMT decreased the mRNA and protein levels of Collagen Ⅰ,Collagen Ⅲ and FN associated with fibrotic phenotype of THESCs (P<0.05),and reduced the phosphorylation level of Smad3 protein,downstream of TGF-β1 (P<0.05).After activation of TGF-β1/Smad3 signaling pathway,the protein levels of TGF-β1/Smad3 signaling pathway and downstream fibrosis phenotype molecules,Collagen Ⅲ and FN,were significantly decreased in the LV-GNMT+SRI-011381 group.Conclusion Overexpression of GNMT can inhibit endometrial fibrosis by regulating TGF-β1/Smad3 signaling pathway,thus achieving therapeutic effect on IUA.

Objective To explore the intrinsic steady-state electrophysiological properties of mouse heart under physiological conditions by high-resolution quantitative analysis.Methods Twenty-two young adult C57BL/6 mice with a 1:1 male-to-female ratio were used.The limbs of the mice were fixed without anesthesia,and electrocardiographic waveforms,including characteristic P-waves,R-waves,and ST-waves,were recorded using a sensitive 12-lead electrophysiological recorder(ECGsqa)under spontaneous breathing.LabScribe software was used to extract and quantify high-resolution time course and amplitude parameters within a single cardiac cycle from the V3 precordial lead.Pearson correlation test combined with simple linear regression was used to generate a scatter plot of ECG parameter fitting.The common and unique correlation parameters were separately identified by joint associations for profiling the quantitative association network.Results ECGsqa analysis identified and quantified 14 characteristic ECG parameters,28.6%of which showed statistical differences between the groups.Compared to male mice,female mice exhibited higher amplitudes and velocities of R and ST waves.Among the 51 association pairs identified in primary association analysis,47.1%were positively correlated,including shared(29.2%),male-specific(29.2%),and female-specific(41.7%)association groups.Second-order clustering of the association pairs revealed that the amplitude-rate association pairs of each waveform voltage in both male and female mouse hearts were strongly correlated.The male mice showed an atrioventricular interconnection pattern,while the female mice showed a unique atrial conduction system quality dependence.The distribution network characteristics of the association groups showed that sex-specific and common correlation sets formed a certain series pattern.Conclusion We discovered a novel intrinsic correlation network of cardiac electrophysiological traits in male and female mice,which reveals the key internal quantitative characteristics and gender difference of both atrial and ventricular conduction systems.

Objective To explore the intrinsic steady-state electrophysiological properties of mouse heart under physiological conditions by high-resolution quantitative analysis.Methods Twenty-two young adult C57BL/6 mice with a 1:1 male-to-female ratio were used.The limbs of the mice were fixed without anesthesia,and electrocardiographic waveforms,including characteristic P-waves,R-waves,and ST-waves,were recorded using a sensitive 12-lead electrophysiological recorder(ECGsqa)under spontaneous breathing.LabScribe software was used to extract and quantify high-resolution time course and amplitude parameters within a single cardiac cycle from the V3 precordial lead.Pearson correlation test combined with simple linear regression was used to generate a scatter plot of ECG parameter fitting.The common and unique correlation parameters were separately identified by joint associations for profiling the quantitative association network.Results ECGsqa analysis identified and quantified 14 characteristic ECG parameters,28.6%of which showed statistical differences between the groups.Compared to male mice,female mice exhibited higher amplitudes and velocities of R and ST waves.Among the 51 association pairs identified in primary association analysis,47.1%were positively correlated,including shared(29.2%),male-specific(29.2%),and female-specific(41.7%)association groups.Second-order clustering of the association pairs revealed that the amplitude-rate association pairs of each waveform voltage in both male and female mouse hearts were strongly correlated.The male mice showed an atrioventricular interconnection pattern,while the female mice showed a unique atrial conduction system quality dependence.The distribution network characteristics of the association groups showed that sex-specific and common correlation sets formed a certain series pattern.Conclusion We discovered a novel intrinsic correlation network of cardiac electrophysiological traits in male and female mice,which reveals the key internal quantitative characteristics and gender difference of both atrial and ventricular conduction systems.

AIM:This study aims to investigate the mechanisms by which Gegen(Puerariae lobatae Radix)and Danshen(Salviae miltiorrhizae Radix et Rhizoma)intervene in diabetes mellitus(DM)-related atherosclerosis(AS)through the regulation of adipokine homeostasis.METHODS:A mouse model of DM-related AS was established in male apolipoprotein E gene knockout(ApoE-/-)mice by high-fat diet feeding and intraperitoneal injection of streptozotocin.The mice were randomly divided into model group,low-dose Gegen-Danshen group,high-dose Gegen-Danshen group and posi-tive control drug metformin group,while male C57BL/J mice served as blank control group,with 6 mice in each group.Se-rum lipid levels of the mice were assessed.Oil red O staining was employed to observe pathological changes in the abdomi-nal aorta,while HE staining was used to examine the morphological changes of adipose tissues surrounding abdominal or-gans.The protein expression levels of adipokines,including adiponectin,apelin,chemerin and resistin,in adipose tis-sues were measured using RT-qPCR and Western blot.RESULTS:The administration of Gegen and Danshen resulted in a significant increase in the protein expression levels of adiponectin and apelin in adipose tissues(P＜0.01).Conversely,the levels of chemerin and resistin were significantly decreased(P＜0.01),and there was a notable reduction in low-densi-ty lipoprotein cholesterol(P＜0.01).Results of oil red O staining revealed that the aortic AS plaques of the mice in model group were numerous,large and scattered,whereas those in high-dose Gegen-Danshen group were fewer and smaller.CONCLUSION:Gegen and Danshen play a protective role in preventing DM-related AS in mice by enhancing the expres-sion of adiponectin and apelin but inhibiting the expression of chemerin and resistin.

AIM:This study aims to investigate the mechanisms by which Gegen(Puerariae lobatae Radix)and Danshen(Salviae miltiorrhizae Radix et Rhizoma)intervene in diabetes mellitus(DM)-related atherosclerosis(AS)through the regulation of adipokine homeostasis.METHODS:A mouse model of DM-related AS was established in male apolipoprotein E gene knockout(ApoE-/-)mice by high-fat diet feeding and intraperitoneal injection of streptozotocin.The mice were randomly divided into model group,low-dose Gegen-Danshen group,high-dose Gegen-Danshen group and posi-tive control drug metformin group,while male C57BL/J mice served as blank control group,with 6 mice in each group.Se-rum lipid levels of the mice were assessed.Oil red O staining was employed to observe pathological changes in the abdomi-nal aorta,while HE staining was used to examine the morphological changes of adipose tissues surrounding abdominal or-gans.The protein expression levels of adipokines,including adiponectin,apelin,chemerin and resistin,in adipose tis-sues were measured using RT-qPCR and Western blot.RESULTS:The administration of Gegen and Danshen resulted in a significant increase in the protein expression levels of adiponectin and apelin in adipose tissues(P＜0.01).Conversely,the levels of chemerin and resistin were significantly decreased(P＜0.01),and there was a notable reduction in low-densi-ty lipoprotein cholesterol(P＜0.01).Results of oil red O staining revealed that the aortic AS plaques of the mice in model group were numerous,large and scattered,whereas those in high-dose Gegen-Danshen group were fewer and smaller.CONCLUSION:Gegen and Danshen play a protective role in preventing DM-related AS in mice by enhancing the expres-sion of adiponectin and apelin but inhibiting the expression of chemerin and resistin.

Objective To investigate the genetic diversity and genetic differentiation of different geographical isolates of Gohieria fusca.. Methods G. fusca isolates were sampled from Wuhu (WH), Bengbu (BB) and Bozhou cities (BZ) of Anhui Province and Jiaxing City of Zhejiang Province (JX). Mitochondrial cytochrome b (Cytb) and ribosomal internal transcribed spacer (ITS) genes were amplified in WH, BB, BZ and JX isolates of G. fusca using PCR assay. The gene sequences were edited and aligned using the software Chromas 2 and DNASTAR 1.00, and the haplotype, haplotype diversity (Hd) and nucleotide polymorphism (Pi) of each isolate were calculated using the software DnaSP 5.10.00. The genetic differentiation among isolates (F_st) and gene flow value (N_m) were estimated using the software MEGA 10.2, and a phylogenetic tree was built. Tests of neutrality and analysis of molecular variance (AMOVA) were performed using the software Arlequin 3.1 and a haplotype network was built based on the Median-Joining network using the software Network 10.2. Results PCR assay showed that the sizes of the Cytb and ITS genes were 372 bp and 1 301 to 1 320 bp, respectively. All four isolates of G. fusca presented high genetic diversity based on mitochondrial Cytb and ITS genes (Hd = 0.804, Pi = 0.006 91). AMOVA showed genetic differentiation among geographical isolates of G. fusca (F_st = 0.202 40, P < 0.05), and the genetic variation was mainly caused by intra-population variations (79.76%). Gene flow analysis showed a high level of gene flow among G. fusca isolates (N_m > 1). Tests of neutrality based on Cytb gene measured a Tajima’s D value of −1.796 31 (P < 0.05) and a Fu’s FS value of −3.293 98 (P < 0.05) in WH isolate of G. fusca, indicating population expansion in WH isolate of G. fusca. Haplotype network analysis and phylogenetic analysis revealed no remarkable geographical distribution pattern among different geographical isolates of G. fusca. All four isolates of G. fusca presented high genetic diversity (Hd = 0.985, Pi = 0.011 97). AMOVA showed moderate level of genetic differentiation between four isolates (F_st = 0.104 62, P < 0.05). The tests of neutrality based on ITS genes measured a Tajima’s D value of −6.088 20 and a Fu’s FS value of −1.935 99 (both P > 0.05) in the whole isolate of G. fusca, indicating no obviously population expansion. Conclusions The four geographical isolates of G. fusca have high genetic diversity and remarkable genetic differentiation. Since a high level of gene flow is detected among different geographical isolates of G. fusca, no obvious geographical distribution pattern of G. fusca is found.

Objective    To compare the outcomes following emergency surgery or conservative treatment for patients with acute type A aortic intramural hematoma (IMH). Methods    Clinical data of consecutive patients diagnosed with acute type A aortic IMH in our hospital from September 2014 to December 2018 were retrospectively analyzed. The patients who met our surgical indications received surgery (an operation group) and other patients received strict conservative treatment (a conservative treatment group). Results    Finally 127 patients were enrolled, including 112 males and 15 females with an average age of 53.6±13.0 years. Of 127 patients, 85 (66.9%) patients accepted emergency surgery and 42 (33.1%) patients accepted strict conservative treatment. There was no difference between the two groups in early mortality or complications (P>0.05). The 5-year survival rate was 90.4% in the operation group and 74.3% in the conservative treatment group (P=0.010). A maximum aortic diameter in the ascending aorta and aortic arch≥45 mm and maximum thickness of IMH in the same section≥8 mm were risk factors for IMH-related death in patients undergoing conservative treatment (P<0.001). Conclusion    The mortality associated with emergency surgery for patients with acute type A aortic IMH is satisfactory. In clinical centers with well-established surgical techniques and postoperative management, emergency surgical treatment may provide a better outcome than conservative treatment for patients with acute type A aortic IMH.

Objective:To identify the causative genes of sporadic thoracic aortic aneurysm or dissection (TAAD) and their correlation with clinical phenotype in the southern Chinese.Methods:We analyzed 11 core genes of TAAD probands without specific family history of 226 cases by next-generation sequencing technology, and performed sanger sequencing for their close relatives. Clinical data of each patient, including age of onset, syndromic phenotypes, involvement of aortic root, aortic maximum diameter and D-dimer were collected. And statistical software SPSS was used to evaluate the correlation between clinical phenotypes and gene mutations.Results:A total of 106 variants were detected in 226 probands with gene-positive frequency of 44.69%, consist of 16 pathogenic (P) variants, 18 likely pathogenic (LP) variants and 71 variants of uncertain significance (VUS). More than half of the mutations were from the non-syndromic TAAD, in which the FBN1 still was the most common causative gene. Earlier age of onset, an increase of women, larger diameter of aorta, Stanford B dissection and severe aortic regurgitation were likely to occur on carriers of P/LP, while thoracic aortic aneurysm occurs on carriers of VUS. Phenotype of both syndrome and dissection with aneurysm could increase the likelihood of carrying gene mutation, but D-dimer and involvement of aortic root couldn’t.Conclusion:Patients with sporadic aortic diseases in southern China have significant genetic heterogeneity and specific correlation between their clinical phenotype and gene mutation, especially in non-syndromic population. Earlier age of onset in carriers of FBN1 or ACTA2 genes, and larger maximum diameters of aorta in carrier of P/LP.